Cytotherapy 23 (2021) 510 520

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FULL-LENGTH ARTICLE

Translational Research

Reversion of early- and late-stage b-cell dedifferentiation by human

umbilical cord-derived mesenchymal stem cells in type 2 diabetic mice
Bing Li1,*, Yu Cheng1,*, Yaqi Yin1, Jing Xue1, Songyan Yu1, Jieqing Gao2, Jiejie Liu3,
Li Zang, PhD1,**, Yiming Mu, PhD1,**
1
Department of Endocrinology, First Medical Center of People’s Liberation Army General Hospital, Beijing, China
2
Department of Endocrinology, Beijing Rehabilitation Hospital of Capital Medical University, Beijing, China
3
Department of Molecular Biology, Institute of Basic Medicine, School of Life Science, People’s Liberation Army General Hospital, Beijing, China

A R T I C L E I N F O A B S T R A C T

Article History: Background aims: The authors aimed to observe b-cell dedifferentiation in type 2 diabetes mellitus (T2DM)
Received 7 July 2020 and investigate the reversal effect of umbilical cord-derived mesenchymal stem cells (UC-MSCs) on early-
Accepted 20 January 2021 and late-stage b-cell dedifferentiation.
Key Words: Methods: In high-fat diet (HFD)/streptozotocin (STZ)-induced T2DM mice, the authors examined the predom-
b-cell dedifferentiation inant role of b-cell dedifferentiation over apoptosis in the development of T2DM and observed the reversion
mesenchymal stem cells of b-cell dedifferentiation by UC-MSCs. Next, the authors used db/db mice to observe the progress of b-cell
type 2 diabetes mellitus dedifferentiation from early to late stage, after which UC-MSC infusions of the same amount were performed
cell transplantation in the early and late stages of dedifferentiation. Improvement in metabolic indices and restoration of b-cell
dedifferentiation markers were examined.
Results: In HFD/STZ-induced T2DM mice, the proportion of b-cell dedifferentiation was much greater than that
of apoptosis, demonstrating that b-cell dedifferentiation was the predominant contributor to T2DM. UC-MSC
infusions significantly improved glucose homeostasis and reversed b-cell dedifferentiation. In db/db mice,
UC-MSC infusions in the early stage significantly improved glucose homeostasis and reversed b-cell dedifferen-
tiation. In the late stage, UC-MSC infusions mildly improved glucose homeostasis and partially reversed b-cell
dedifferentiation. Combining with other studies, the authors found that the reversal effect of UC-MSCs on
b-cell dedifferentiation relied on the simultaneous relief of glucose and lipid metabolic disorders.
Conclusions: UC-MSC therapy is a promising strategy for reversing b-cell dedifferentiation in T2DM, and the
reversal effect is greater in the early stage than in the late stage of b-cell dedifferentiation.
© 2021 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.

Introduction development of T2DM [4]. Although the progression of b-cell dys-

The incidence of diabetes has seen a dramatic increase in the past
decades, and diabetes has become an epidemic worldwide [1]. It is
estimated that global diabetes prevalence will rise to 10.2% by 2030
and 10.9% by 2045 [2]. Type 2 diabetes mellitus (T2DM), constituting
over 90% of all diabetes cases, is regarded as the major contributor to
the total diabetes population, and research on effective prevention
and treatment of T2DM will have great benefits [3]. Progressive
b-cell dysfunction—namely, decrease in insulin biosynthesis and
secretion and loss of b-cell numbers—plays a central role in the
** Correspondence: Li Zang and Yiming Mu, Department of Endocrinology, The First
Medical Center of PLA General Hospital, Beijing 100853, China.
* These authors contributed equally to this work.
E-mail addresses: lzang301@163.com (L. Zang), muyiming@301hospital.com.cn
(Y. Mu).
function may be ascribed to b-cell apoptosis, recent studies have sug-
gested that increased apoptotic b cells do not proportionally
correlate with the islet functional impairment in T2DM [5]. Moreover,
the fact that b-cell dysfunction can be partly and temporarily
restored by dietary or pharmacological interventions indicates that
the loss of b-cell numbers or function is not permanent, corroborat-
ing the view that apoptosis is not the dominant cause of b-cell dys-
function in T2DM [6,7]. Strikingly, increasing studies have put
forward that b-cell dysfunction is mainly due to b-cell dedifferentia-
tion (loss of canonical b-cell markers and regression to an endocrine
progenitor-like stage) in diabetes models [8-10] as well as in human
T2DM [11]. Commonly used hypoglycemic drugs act by enhancing
insulin sensitivity in peripheral tissues (e.g., thiazolidinediones), by
suppressing hepatic gluconeogenesis (e.g., biguanides), by increasing
insulin secretion in a glucose-dependent fashion (e.g., glucagon-like

https://doi.org/10.1016/j.jcyt.2021.01.005
1465-3249/© 2021 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.
 B. Li et al. / Cytotherapy 23 (2021) 510 520
peptide 1 analogues) and by boosting glucose elimination via the kid-
neys (e.g., sodium-glucose cotransporter 2 inhibitors). Unexpectedly,
results from Ishida et al. [12] showed that hypoglycemic drugs,
including the insulin-sensitizer rosiglitazone, sodium-glucose
cotransporter 2 inhibitor phloridzin and insulin, could lower blood
glucose levels but could not reverse b-cell dedifferentiation.
Mesenchymal stem cells (MSCs) are a heterogeneous subset of stro-
mal stem cells that can be isolated from various adult tissues, and they
possess self-renewal capacity, multi-lineage differentiation potential,
paracrine effects and immunomodulatory properties, which makes
them a promising tool in regenerative medicine. Human umbilical
cord-derived MSCs (UC-MSCs) display higher proliferative potential and
lower immunogenicity with allogeneic sources than other commonly
used MSCs, such as bone marrow-derived MSCs (BM-MSCs). In recent
years, UC-MSC therapy has made remarkable achievements in treating
T2DM. A clinical study in T2DM patients demonstrated that two infu-
sions of UC-MSCs significantly lowered blood glucose and glycosylated
hemoglobin, improved islet function and decreased the incidence of
diabetic complications during a follow-up period of 36 months [13].
The authors’ group has long been committed to experimental research
concerning UC-MSC therapy in T2DM. One of the authors’ previous
studies demonstrated that UC-MSCs could exert anti-diabetic effects by
regulating macrophage phenotype and alleviating insulin resistance in
high-fat diet (HFD)/streptozotocin (STZ)-induced T2DM rats [14]. How-
ever, the mechanism by which UC-MSCs attenuate b-cell dysfunction
has not been fully elucidated. Another one of the authors’ studies on
HFD/STZ-induced T2DM mice revealed that UC-MSC infusion sup-
pressed inflammation in the islet microenvironment, thereby leading to
increased insulin+ b cells and improved islet function. However, the
promoted proliferation and inhibited apoptosis of b cells after UC-MSC
treatment constituted only a small fraction of the significant increase in
insulin+ b cells. The “origin” of most of the increased b cells remained
elusive. The authors speculated that the increased b cells derived
largely from redifferentiation and maturation of dedifferentiated b cells
by UC-MSCs, and the mechanistic action of UC-MSCs in mitigating
b-cell dysfunction might be the reversion of b-cell dedifferentiation in
T2DM.
Recently, Wang et al. [15] inspiringly showed in 7- to 9-week-old db/
db mice that MSCs attenuated b-cell dysfunction by reversing b-cell
dedifferentiation, suggesting that the increased b cells may be rediffer-
entiated b cells. However, the db/db mice used in this study were in the
early stage of b-cell dedifferentiation; the effect of MSCs on reversing
late-stage b-cell dedifferentiation was not investigated. In the present
study, the authors further tracked the characteristic trends of b-cell
dedifferentiation in the natural history of T2DM in both HFD/STZ-
induced T2DM mice and db/db mice, observed the transition from early-
to late-stage b-cell dedifferentiation (marked by the emergence of neu-
rogenin 3+ [Ngn3+] cells in the islets) at a series of time points and inves-
tigated the reversal effect of UC-MSCs on both early- and late-stage
b-cell dedifferentiation. The results showed that UC-MSCs effectively
curtailed markers of b-cell dedifferentiation in both T2DM models; in
db/db mice, UC-MSCs reversed early-stage b-cell dedifferentiation better
than late-stage b-cell dedifferentiation. Moreover, the reversal effect of
UC-MSCs on b-cell dedifferentiation in db/db mice relied on the simulta-
neous improvement in glucose and lipid homeostasis. The authors’ study
further highlighted UC-MSCs’ potential in ameliorating b-cell dysfunc-
tion by targeting b-cell dedifferentiation, a dominant mechanism in
T2DM, and provided an experimental basis for choosing an appropriate
time for UC-MSC treatment in clinical applications.
Methods
Isolation, culture and characterization of human UC-MSCs
Human umbilical cords were obtained from women giving birth
in the First Medical Center of Chinese People’s Liberation Army
511
General Hospital. All subjects provided informed consent. Ethics
approval was obtained from the Ethics Committee of the Chinese PLA
General Hospital. The experimental protocols were approved by the
Medical Ethics Committee of People’s Liberation Army General Hos-
pital. Human UC-MSCs were isolated and cultured as per previously
described methods [16]. The cultured UC-MSCs at passage three were
digested and harvested to identify the immunophenotype using fluo-
rescein isothiocyanate-conjugated anti-CD45, anti-CD90 and anti-
HLA-DR; phycoerythrin-conjugated anti-CD105 and anti-CD73; and
peridinin-chlorophyll-protein complex-conjugated anti-CD34 anti-
bodies using flow cytometry analysis. All antibodies used for surface
marker analysis were purchased from BD Biosciences (San Diego, CA,
USA). The potential of UC-MSCs to differentiate into osteoblasts and
adipocytes was verified as per previously described methods [17].
Animals and induction of T2DM model
Two types of T2DM mouse models were used in the present study.
The first model was an HFD/STZ-induced T2DM model. Eight-week-old
male C57BL/6J mice, purchased from the First Medical Center of Chinese
People’s Liberation Army General Hospital, were raised in a tempera-
ture- and humidity-controlled environment under a 12-h light/12-h
dark cycle. T2DM was established via a combination of 12-week HFD
and intraperitoneal injection of low-dose STZ (100 mg/kg, Sigma-
Aldrich, St. Louis, MO) as per previously described methods [18]. One
week after STZ injection, intraperitoneal glucose tolerance tests (IPGTTs)
and intraperitoneal insulin tolerance tests (IPITTs) were conducted to
confirm the T2DM model. For IPGTT and IPITT, mice were starved over-
night and intraperitoneally injected with glucose (1.5 g/kg) or insulin (1
U/kg). Blood glucose levels, determined by tail blood samples, were
measured every 30 min up to 120 min. During the observation period,
no death or obvious toxic reaction occurred. The second model was
genetically obese, leptin receptor-deficient db/db mice, a spontaneous
T2DM model. Eight-week-old male C57BL/6 db/db mice and age-
matched C57BL/6 wild-type db/+ littermates, originally obtained from
The Jackson Laboratory, were provided by the Peking University Health
Science Center and fed a standard chow diet in a temperature- and
humidity-controlled environment under a 12-h light/12-h dark cycle.
Experimental design
In the HFD/STZ-induced T2DM model, the authors first observed
the trends of b-cell dedifferentiation and apoptosis within the 4
weeks of T2DM. Weight and blood glucose in normal and T2DM mice
were monitored weekly. At 1 week, 2 weeks, 3 weeks, 4 weeks and 5
weeks after intraperitoneal injection of STZ, a random selection of
T2DM mice (n = 6) and normal mice (n = 6) were killed, and the peak
time point of b-cell dedifferentiation was determined by immunoflu-
orescence analysis of the pancreata. Next, passage three UC-MSCs
were used to investigate whether UC-MSCs could reverse b-cell
dedifferentiation. UC-MSC infusions (1 £ 106 UC-MSCs suspended in
200 mL phosphate-buffered saline [PBS]) into T2DM mice (n = 6) via
the tail vein were performed once a week for 2 weeks, starting from
the peak time point of b-cell dedifferentiation. The control T2DM
mice (n = 6) were infused with 200 mL PBS at the same time. The nor-
mal C57BL/6J mice (n = 6) received no treatment. Weight and blood
glucose in each group were monitored weekly. One week after UC-
MSC infusions, IPGTTs and IPITTs were conducted to evaluate the
therapeutic effects of UC-MSCs. After these tests, mice in each group
were killed for immunofluorescence analysis of the pancreata to
examine the expression of b-cell dedifferentiation-related markers.
In db/db mice, a widely used model for studying human T2DM,
the authors first observed the trends and staged characteristics of
b-cell dedifferentiation in 10- to 32-week-old mice, with age-
matched wild-type db/+ mice used as controls. Weight and blood glu-
cose were monitored weekly, and a random selection of db/db mice
512 B. Li et al. / Cytotherapy 23 (2021) 510 520
(n = 6) and db/+ mice (n = 6) were killed at 12, 16, 20, 23, 26 and 30
weeks of age. Immunofluorescence analysis of the pancreata at the
indicated time points was performed to determine the time points
(represented as age of the mice) of early- and late-stage b-cell dedif-
ferentiation. Next, UC-MSCs (1 £ 106 UC-MSCs suspended in 200 mL
PBS) were infused through the tail vein into db/db mice starting from
the early or late stage of b-cell dedifferentiation once a week for 6
weeks. The age-matched control db/db mice were infused with 200
mL PBS at the same time. Weight and blood glucose in each group
(n = 6) were monitored weekly. One week after UC-MSC infusions,
IPGTTs and IPITTs were conducted to evaluate the therapeutic effects
of UC-MSCs in the early and late treatment groups. After these tests,
mice in each group were killed for immunofluorescence analysis of
the pancreata to examine the expression of b-cell dedifferentiation-
related markers.
Blood and tissue collection
For the HFD/STZ-induced T2DM model, mice were first injected
intraperitoneally with 1% pentobarbital sodium (50 mg/kg) anesthe-
sia at the indicated time points at which they were killed. Thereafter,
mice were perfused through the left ventricle with 10 15 mL PBS,
followed by 10 15 mL of 4% paraformaldehyde. After perfusion, the
pancreas was collected, incubated in 30% sucrose/phosphate buffer
overnight and then embedded in Tissue-Tek optimal cutting temper-
ature compound (Sakura Finetek, Torrance, CA, USA) and cut into fro-
zen sections 5-mm-thick for immunofluorescence staining.
The db/db mice were first anesthetized at the indicated time
points, after which blood was collected and centrifuged at 3000 rpm
for 15 min to obtain serum for biochemical analyses. Mice were then
successively perfused through the left ventricle with PBS and parafor-
maldehyde, and the pancreas was collected and embedded for immu-
nofluorescence staining as just described. The levels of total
cholesterol (TC), triglyceride (TG) and low-density lipoprotein choles-
terol (LDL-C) in the serum were measured using standard analytics
(Kang Jia Hong Yuan Biological Technology Co Ltd, Beijing, China).
Immunofluorescence staining
For immunofluorescence staining, the frozen sections were incu-
bated for 14 h at 4°C with primary antibodies specific for insulin at
1:100 (ab7842; Abcam, Cambridge, UK), Pdx1 at 1:400 (5679; CST,
Boston, MA, USA), Foxo1 at 1:100 (2880; CST), ALDH1A3 at 1:100
(NBP2-15339; Novus-biologicals, Littleton, CO), chromogranin A
(ChrA) at 1:200 (ER40724; HuaBio), Ngn3+ at 1:200 (AB5684; Milli-
pore) or terminal deoxynucleotidyl transferase dUTP nick end label-
ing (TUNEL) (Roche, Mannheim, Germany), then with Alexa Fluor
488/594-conjugated secondary antibodies at 1:500 (Invitrogen, NY,
USA) for 2 h at room temperature. The nuclei were visualized with
40 ,6-diamidino-2-phenylindole (Sigma-Aldrich, St Louis, MO, USA).
The immunofluorescence-stained sections were captured using a
laser scanning confocal microscope (Leica, Wetzlar, Germany). Islet
cells double-stained by ChrA/insulin, Pdx1/insulin, Foxo1/insulin and
ALDH1A3/insulin were counted. The mean percentage of ChrA+/insu-
lin+ cells was calculated as average ChrA+/insulin+ cell number/total
ChrA+ cell number in the islet £ 100% to examine the proportion of
insulin-producing b cells. The mean percentage of Pdx1+/insulin+ or
Foxo1+/insulin+ cells was calculated as average Pdx1+/insulin+ or
Foxo1+/insulin+ cell number/total islet cell number £ 100% to evalu-
ate the expression of Pdx1 and Foxo1. In addition, ALDH1A3-positive
cells were counted, and the mean percentage of ALDH1A3+ cells was
calculated as average ALDH1A3+ cell number/total islet cell
number £ 100% to examine the proportion of dedifferentiated b cells
in the islet. TUNEL+ and Ngn3+ cells were also counted, and the
results were presented as the number of positive cells per islet. At
least three random microscopic fields per sample and five random
islets per microscopic field were scored.
Statistical analysis
The results were expressed as mean § standard deviation. The sta-
tistical analysis was carried out using SPSS Statistics 19.0 software (IBM,
Armonk, NY, USA). Differences between means were assessed using
t-tests (two samples) or one-way ANOVA (three or more samples). For
all analyses, the statistical significance level was set at P < 0.05.
Results
HFD/STZ-treated mice developed hyperglycemia with impaired islet
function, decreased insulin sensitivity and loss of b-cell identity
Successful establishment of T2DM was confirmed by monitoring
blood glucose, weight, IPGTTs and IPITTs. Before STZ injection, the
mice fed with HFD developed obesity, and body weight measured
47.1 § 1.0 g, which was markedly higher than that of mice fed normal
chow (32.0 § 0.6 g). One week after STZ injection, the blood glucose
in STZ-treated mice rose to and fluctuated between 20 mmol/L and
30 mmol/L (see supplementary Figure 1A), whereas body weight in
STZ-treated mice decreased significantly (see supplementary Figure
1B). IPGTT results indicated significantly impaired glucose metabo-
lism, and IPITT results showed markedly deteriorated insulin sensi-
tivity in HFD/STZ-induced mice, thereby indicating the successful
establishment of T2DM (see supplementary Figure 1C,D). Immunoflu-
orescence analysis of the pancreata was conducted at the indicated
time points to observe morphological changes in pancreatic islets
and examine expression of ChrA (a general endocrine marker) as
well as Pdx1 and Foxo1 (two key b-cell-enriched transcription fac-
tors) and ALDH1A3 (a newly defined marker of b-cell dedifferentia-
tion) in the islets. In addition, TUNEL staining was performed to
examine b-cell apoptosis. ChrA/insulin double immunofluorescence
staining (Figure 1A) showed morphological destruction and signifi-
cant reduction in ChrA+/insulin+ b cells in the islets of T2DM mice 1
week after STZ injection compared with normal mice. Starting from 1
week to 5 weeks after STZ injection, the percentage of ChrA+/insulin+
cells progressively decreased from 57.5 § 4.8% to 29.6 § 2.1%
(Figure 1F), and the percentage of Pdx1+/insulin+ cells and Foxo1+/
insulin+ cells in the islets gradually declined from 46.7 § 7.2% and
49.7 § 2.1% to 4.9 § 2.2% and 5.0 § 0.9%, respectively (Figure 1B,C,G,
H), thereby indicating the reduction in functional b cells and loss of
b-cell identity in the development T2DM. Moreover, the percentage
of ALDH1A3+ cells in the islets of T2DM mice significantly increased
compared with normal mice, reaching the highest level (15.7 § 2.1%)
at 3 weeks after STZ injection (Figure 1D,I). Additionally, TUNEL/insu-
lin double staining (Figure 1E) showed that TUNEL+ b cells rarely
existed in normal mice. After STZ injection, an average of one TUNEL+
apoptotic b cell per islet (0.82 § 0.31%) was detected at the indicated
time points. Based on these results, the authors believed that dedif-
ferentiation instead of apoptosis was the dominant contributor to
b-cell “loss” and islet dysfunction in T2DM. Moreover, Ngn3+, an islet
progenitor marker, was not detected during the observation period
(see supplementary Figure 2), suggesting that b cells were in the
early stage of dedifferentiation within the first 4 weeks of T2DM in
HFD/STZ-induced mice.
UC-MSC infusions reversed b-cell dedifferentiation in HFD/STZ-induced
T2DM mice
According to the aforementioned results, b-cell dedifferentiation
predominated over apoptosis as a mechanism of b-cell dysfunction in
HFD/STZ-induced T2DM mice. Using ALDH1A3, a marker of b-cell dedif-
ferentiation, the authors found that T2DM mice displayed augmented
 B. Li et al. / Cytotherapy 23 (2021) 510 520 513

Fig. 1. Analysis of b-cell dedifferentiation and apoptosis during the development of T2DM in HFD/STZ-induced type 2 diabetic mice. Mice in normal and T2DM groups were killed at
1 week, 2 weeks, 3 weeks, 4 weeks and 5 weeks after STZ injection. (A D) Immunofluorescence staining of ChrA (red), Pdx1 (red), Foxo1 (red), ALDH1A3 (red) and insulin (green) was
performed. Representative images of ChrA/insulin, Pdx1/insulin, Foxo1/insulin and ALDH1A3/insulin, with arrows indicating the Pdx1+/insulin+ cell, Foxo1+/insulin+ cell and ALDH1A3+
cell in the amplified image. (E) Apoptosis of insulin-producing cells (red) was evaluated by TUNEL labeling (green), with the arrow showing the insulin+ cells expressing TUNEL. Nuclei
were labeled with DAPI. The percentage of insulin+ cells co-expressing ChrA (F), Pdx1 (G) and FOXO1 (H) and the percentage of ALDH1A3+ cells (I) per islet were quantified. Scale
bar = 50 mm. Data are expressed as mean § SD. **P < 0.01. DAPI, 40 ,6-diamidino-2-phenylindole; SD, standard deviation; w, week(s). (Color version of figure is available online).

islet expression of ALDH1A3 that reached the highest level at 3 weeks
after STZ injection. Next, T2DM mice were randomly assigned to the
T2DM group (treated with PBS infusions) or the UC-MSC group (treated
with UC-MSC infusions). Mice fed standard chow were used as normal
controls. Starting from the third week after STZ injection, T2DM mice
were infused with PBS/UC-MSCs via the tail vein once a week for 2
weeks. The results (Figure 2A) showed that the T2DM group remained
severely hyperglycemic (28.2 § 3.2 mmol/L) during the observation
period. After the first infusion, the UC-MSC group showed a gradual
decrease in random blood glucose (25.2 § 2.6 mmol/L), and after the
second infusion, blood glucose in the UC-MSC group further dropped to
22.6 § 1.9 mmol/L, which was significantly lower than that observed in
the T2DM group. Mice in the T2DM and UC-MSC groups experienced
weight loss after STZ injection, and UC-MSC infusions did not affect
body weight between the two groups (Figure 2B). The IPGTTs
(Figure 2C) and IPITTs (Figure 2D) showed that, compared with the
T2DM group, the UC-MSC group had evidently alleviated glucose intol-
erance and increased insulin sensitivity. These results demonstrated
that UC-MSC infusions significantly improved glucose homeostasis in
HFD/STZ-induced T2DM mice.
To further investigate the efficacy of UC-MSCs on b-cell dediffer-
entiation, immunofluorescence staining on pancreata was performed.
Overall, the results showed that UC-MSC infusions significantly
ameliorated islet damage, including morphological disorganization
and reduction in islet size. A significant restoration of functional b
cells per islet was evidenced by a dramatic increase in the percentage
of ChrA+/insulin+ cells from 31.7 § 2.4% to 63.1 § 1.2% (Figure 2E),
with Pdx1+/insulin+ cells increasing from 9.5 § 1.4% to 43.9 § 2.3%
(Figure 2F) and Foxo1+/insulin+ cells increasing from 15.6 § 1.3% to
31.1 § 2.1% (Figure 2G). Moreover, UC-MSC infusions resulted in a
significant reduction in ALDH1A3+ cells (from 15.1 § 1.4% to 6.4 §
2.2%) (Figure 2H) in the islets. Taken together, these results suggested
that UC-MSC infusions exerted remarkable anti-diabetic effects and
curtailed the markers of b-cell dedifferentiation in HFD/STZ-induced
T2DM mice. In summary, insulin+ b-cell numbers in the islets of
MSC-treated mice increased by “benefitting” from the reversal effect
of UC-MSCs on b-cell dedifferentiation.
Hyperglycemia drove dedifferentiation of pancreatic b-cell to Ngn3+
precursor-like phenotypes in db/db mice
Db/db mice capture key features of human T2DM, such as obesity,
insulin resistance, hyperglycemia and dyslipidemia. It has been
reported that db/db mice display noticeable markers of b-cell dedif-
ferentiation with the development of T2DM [11,19]. Therefore, db/db
mice and age-matched db/+ mice were used to further explore the
514 B. Li et al. / Cytotherapy 23 (2021) 510 520

Fig. 2. UC-MSC infusion reversed b-cell dedifferentiation in HFD/STZ-induced T2DM mice. Random blood glucose (A) and body weight (B) of the indicated groups were monitored
weekly after STZ injection. Two weeks after the establishment of T2DM (3 weeks after STZ injection), T2DM mice were administered UC-MSC/PBS infusion once a week for 2 weeks.
One week after UC-MSC infusion, glucose tolerance was assessed by IPGTT (C) and insulin sensitivity was assessed by IPITT (D). For IPITT, the result of each time point is presented
relative to initial blood glucose. (E H) Immunofluorescence staining of ChrA (red), Pdx1 (red), Foxo1 (red), ALDH1A3 (red) and insulin (green) was performed. Representative
images of ChrA/insulin, Pdx1/insulin, Foxo1/insulin and ALDH1A3/insulin in the islet. Nuclei were labeled with DAPI. The percentage of insulin+ cells co-expressing ChrA, Pdx1 and
FOXO1 and the percentage of ALDH1A3+ cells in the islet were quantified. Scale bar = 100 mm. Data are expressed as mean § SD. *P < 0.05, **P < 0.01. DAPI, 40 ,6-diamidino-2-phe-
nylindole; ns, not significant ( P > 0.05); SD, standard deviation. (Color version of figure is available online).

characteristics of b-cell dedifferentiation and investigate the reversal
effect of UC-MSCs on b-cell dedifferentiation. As shown in Figure 3A,
B, 10-week-old db/db mice developed distinctive hyperglycemia and
obesity, with random blood glucose rising to 23.7 § 5.3 mmol/L and
body weight reaching 47.7 § 1.9 g. With increasing age, db/db mice
demonstrated gradually elevated blood glucose and body weight. In
20-week-old db/db mice, blood glucose rose to 32.0 § 2.2 mmol/L
and body weight reached 61.5 § 4.0 g. Starting from 20 weeks, blood
glucose in most db/db mice exceeded the maximum value (33.3
mmol/L) that could be detected by the glucometer (those data were
uniformly recorded as 33.3 mmol/L), whereas body weight was rela-
tively stable. Throughout the observation period, blood glucose and
body weight of db/db mice were significantly higher than those of
age-matched db/+ mice.
The dedifferentiation stage of b cells was probed by immunofluo-
rescence analysis of Pdx1, ALDH1A3 and Ngn3+ expression in the
islets of db/db mice. Since Ishida et al. [12] provided evidence that b
cells in 16- to 20-week-old db/db mice appeared to be in the early
stage of dedifferentiation, the authors first examined the expression
of Pdx1 and ALDH1A3 in mice younger than 20 weeks old. As shown
in Figure 3C,D, the percentage of Pdx1+/insulin+ cells significantly
decreased from 79.2 § 1.7% to 58.4 § 1.7%, whereas the percentage
of ALDH1A3+ cells significantly increased from 1.4 § 0.4% to 40.4 §
3.2% in 12-week-old db/db mice, indicating that the b cells had
already begun to dedifferentiate at this time point. With the progress
of T2DM, the percentage of Pdx1+/insulin+ cells in db/db mice further
dropped to 48.9 § 1.7% in 16 weeks and to 19.1 § 4.6% in 20 weeks,
which was significantly lower than that of age-matched db/+ mice. In
addition, the percentage of ALDH1A3+ cells remained high in 16- and
20-week-old db/db mice. Since Talchai et al. [8] provided evidence
that in persistent hyperglycemia, b cells could regress to an Ngn3+
endocrine progenitor phenotype, which was considered the late
stage of b-cell dedifferentiation, the authors then tracked the expres-
sion of Ngn3+ in the islets of db/db mice from 23 weeks onward. As
 B. Li et al. / Cytotherapy 23 (2021) 510 520 515

Fig. 3. Analysis of b-cell dedifferentiation in db/db mice. From 10 weeks on, random blood glucose (A) and body weight (B) were monitored every 2 weeks in db/db mice and age-
matched db/+ mice. (C,D) Immunofluorescence staining of Pdx1 (red), ALDH1A3 (red) and insulin (green) was performed. Representative images of Pdx1/insulin co-staining and
ALDH1A3/insulin co-staining in the islets of db/+ mice and 12-, 16- and 20-week-old db/db mice. The percentage of Pdx1+/insulin+ cells and the percentage of ALDH1A3+ cells were
quantified. Immunofluorescence staining of Ngn3+ (red) and insulin (green) was performed. Scale bar = 100 mm (C) and 50 mm (D). (E) Representative images of Ngn3+/insulin
co-staining in the islets of db/+ mice and 23-, 26- and 30-week-old db/db mice. The number of Ngn3+ cells per islet was counted, with the arrow indicating Ngn3+ cells in the
islet. Nuclei were labeled with DAPI. Scale bar = 50 mm. Data are expressed as mean § SD. **P < 0.01. DAPI, 40 ,6-diamidino-2-phenylindole;SD, standard deviation; w, week(s).
(Color version of figure is available online).

shown in Figure 3E, Ngn3+ immunoreactivity was hardly detectable
in the islets of db/+ mice as well as 23- and 26-week-old db/db mice.
Notably, a stark increase in Ngn3+ immunoreactivity was detected in
the islets of 30-week-old db/db mice. Ngn3+/insulin co-staining
showed that cells expressing Ngn3+ contained no insulin. Taken
together, these results suggested that in 12-week-old db/db mice,
the b cells began to lose their mature phenotype and underwent
early-stage dedifferentiation, whereas in 30-week-old db/db mice,
the b cells regressed to the Ngn3+ endocrine precursor-like state,
entering the late stage of dedifferentiation. Accordingly, two treat-
ment groups were set up to investigate the reversal effect of UC-
MSCs on early- and late-stage b-cell dedifferentiation; one was the
early treatment group and the other was the late treatment group.
The same dose of UC-MSCs was administered to db/db mice starting
at 12 weeks in the former group and 30 weeks in the latter group.
UC-MSC infusions reversed early-stage b-cell dedifferentiation in db/db
mice
In the early treatment group, db/db mice were administered UC-
MSCs (1 £ 106 UC-MSCs suspended in 200 mL PBS) or an equal volume
of PBS as a control treatment once a week for 6 weeks starting at 12
weeks. As shown in Figure 4A, compared with the control group, the
MSC-treated group exhibited significantly lower random blood glu-
cose levels (24.5 § 1.7 mmol/L) after two infusions (P < 0.05). After six
infusions, blood glucose in the MSC-treated group further decreased to
21.6 § 2.1 mmol/L, whereas blood glucose in the control group
remained 33.3 mmol/L. However, body weight showed no marked
difference between the two groups (Figure 4B). IPGTT results
(Figure 4C) showed that in a fasting state, 90 min and 120 min after
glucose injection, blood glucose in the MSC-treated group was signifi-
cantly lower than that observed in the control group, thereby indicat-
ing a marked improvement in glucose tolerance, whereas IPITT results
(Figure 4D) showed a notable increase in insulin sensitivity after UC-
MSC treatment. The aforementioned results indicated that UC-MSC
infusions at the early stage of b-cell dedifferentiation significantly
improved glucose homeostasis in db/db mice. Furthermore, the plasma
lipid profiles of MSC-treated mice were improved, as manifested by
moderately decreased TC (from 1.83 § 0.08 mmol/L to 1.67 § 0.02
mmol/L) (Figure 4E) and LDL-C (from 0.37 § 0.09 mmol/L to 0.25 §
0.01 mmol/L) (Figure 4F) and significantly decreased TG (from 0.42 §
0.03 mmol/L to 0.22 § 0.02 mmol/L) (Figure 4G). The aforementioned
516 B. Li et al. / Cytotherapy 23 (2021) 510 520

Fig. 4. UC-MSC infusions reversed early-stage b-cell dedifferentiation in db/db mice. Random blood glucose (A) and body weight (B) were monitored weekly in db/db mice and age-
matched db/+ mice beginning at 10 weeks. From 12 weeks on, db/db mice were administered UC-MSC/PBS infusion once a week for 6 weeks. One week after UC-MSC infusions, glu-
cose tolerance was assessed by IPGTT (C) and insulin sensitivity was assessed by IPITT (D). Caret (^) indicates that the blood glucose level exceeded the maximum (33.3 mmol/L)
glucometer reading. For IPITT, the result of each time point is presented relative to initial blood glucose. Serum concentrations of TC (E), LDL-C (F) and TG (G) in the indicated groups
were measured. (H J) Immunofluorescence staining of ChrA (red), Pdx1 (red), ALDH1A3 (red) and insulin (green) was performed. Representative images of ChrA/insulin co-stain-
ing, Pdx1/insulin co-staining and ALDH1A3/insulin co-staining in the islets. Nuclei were labeled with DAPI. The percentage of insulin+ cells co-expressing ChrA and Pdx1 and the
percentage of ALDH1A3+ cells were quantified. Scale bar = 50 mm. Data are expressed as mean § SD. *P < 0.05, **P < 0.01. DAPI, 40 ,6-diamidino-2-phenylindole; ns, not significant
(P > 0.05); SD, standard deviation. (Color version of figure is available online).

results demonstrated that UC-MSCs significantly improved glucose
and lipid homeostasis in the early treatment group.
Next, the authors performed immunofluorescence analysis of
mice pancreata to examine the reversal effect of UC-MSCs on early-
stage dedifferentiation. As shown in Figure 4F,G, a significantly
increased percentage of ChrA+/insulin+ cells (from 26.9 § 2.1% to 49.1
§ 3.3%) and Pdx1+/insulin+ cells (from 29.3 § 1.8% to 41.7 § 2.9%)
and a markedly decreased percentage of ALDH1A3+cells (from 30.3 §
4.5% to 7.1 § 1.6%) were evidenced in the MSC-treated group com-
pared with the control group, suggesting that UC-MSC infusions
reversed early-stage b-cell dedifferentiation.
UC-MSC infusions partially reversed late-stage b-cell dedifferentiation in
db/db mice
In the late treatment group, db/db mice were administered the
same dose of UC-MSCs as the early treatment group starting at 30
weeks. As shown in Figure 5A, the MSC-treated db/db mice demon-
strated a gradual decrease in random blood glucose that decreased to
23.8 § 2.5 mmol/L 1 week after the sixth infusion, which was
significantly lower than that seen in the control group (31.9 § 0.8
mmol/L). Similar to the early treatment group, UC-MSC infusions in the
late treatment group did not significantly reduce body weight in db/db
mice (Figure 5B). IPGTT results (Figure 5C) showed that during the fast-
ing state, blood glucose in the MSC-treated group was significantly
lower than that seen in the control group. After glucose injection, blood
glucose in control db/db mice remained 33.3 mmol/L until 120 min,
whereas blood glucose in the MSC-treated group dropped mildly at
120 min, but the change was not statistically significant. IPITT results
(Figure 5D) showed slightly alleviated insulin resistance in the MSC-
treated group compared with the control group. The plasma lipid pro-
files of MSC-treated mice were also improved, as indicated by moder-
ately decreased TC (from 2.08 § 0.18 mmol/L to 1.83 § 0.06 mmol/L)
(Figure 5E) and LDL-C (from 0.43 § 0.01 mmol/L to 0.32 § 0.04 mmol/
L) (Figure 5F) and significantly decreased TG (from 0.44 § 0.03 mmol/L
to 0.28 § 0.03 mmol/L) (Figure 5G). The aforementioned results demon-
strated that UC-MSC treatment also improved glucose and lipid homeo-
stasis in the late treatment group. Comparing the metabolic parameters
in the early and late treatment groups, the authors found that the
improvement in random glucose, glucose tolerance and insulin
 B. Li et al. / Cytotherapy 23 (2021) 510 520 517

Fig. 5. UC-MSC infusions partially reversed late-stage b-cell dedifferentiation in db/db mice. Random blood glucose (A) and body weight (B) were monitored weekly in db/db mice
and age-matched db/+ mice beginning at 28 weeks. From 30 weeks on, db/db mice were administered UC-MSC/PBS infusion once a week for 6 weeks. One week after UC-MSC infu-
sions, glucose tolerance was assessed by IPGTT (C) and insulin sensitivity was assessed by IPITT (D). Caret (^) indicates that the blood glucose level exceeded the maximum (33.3
mmol/L) glucometer reading. For IPITT, the result of each time point is presented relative to initial blood glucose. Serum concentrations of TC (E), LDL-C (F) and TG (G) in the indi-
cated groups were measured. (H K) Immunofluorescence staining of ChrA (red), Pdx1 (red), ALDH1A3 (red), Ngn3+ (red) and insulin (green) was performed. Representative images
of ChrA/insulin co-staining, Pdx1/insulin co-staining, ALDH1A3/insulin co-staining and Ngn3+/insulin co-staining in the islets. The percentage of insulin+ cells co-expressing ChrA
and Pdx1 and the percentage of ALDH1A3+ cells were quantified. The number of Ngn3+ cells per islet was counted. Scale bar = 50 mm. Data are expressed as mean § SD. *P < 0.05,
**P < 0.01. ns, not significant (P > 0.05); SD, standard deviation. (Color version of figure is available online).

sensitivity was greater in the early treatment group than the late treat-
ment group, whereas the improvement in lipid profiles was similar
between the two groups.
Additionally, immunofluorescence analysis of pancreata demon-
strated a significantly increased percentage of ChrA+/insulin+ cells
from 27.4 § 0.3% to 42.1 § 2.2% (Figure 5H) and markedly reduced
Ngn3+ cells in the islets from 6.0 § 0.7 cells/islet to 0.3 § 0.3 cells/islet
(Figure 5K) after UC-MSC infusions. The percentage of Pdx1+/insulin+
cells showed a significant increase from 19.2 § 3.0% to 30.5 § 3.1%,
whereas the percentage of ALDH1A3+ cells showed a slight decrease
from 52.3 § 0.8% to 46.5 § 2.6%. These results suggested that UC-
MSC infusions partially reversed late-stage b-cell dedifferentiation.
Comparing the results of immunofluorescence staining in the early
and late treatment groups, the authors found that the restoration of
dedifferentiation markers by the same dose of UC-MSCs was better in
the early treatment group than the late treatment group.
Discussion
In the current study, the authors observed the progress of b-cell
dedifferentiation in both HFD/STZ-induced T2DM mice and db/db
mice and found that UC-MSC infusions curtailed markers of b-cell
dedifferentiation; increased b cells in the islets of MSC-treated mice
derived from the redifferentiation of b cells mediated by UC-MSCs.
Moreover, the authors found for the first time that in db/db mice at
30 weeks old, b cells dedifferentiated to a pancreatic progenitor cell
state marked by Ngn3+ expression, indicating that b-cell dedifferenti-
ation turned from early to late stage. UC-MSC infusions in the early
stage significantly reversed b-cell dedifferentiation, whereas infu-
sions in the late stage only partially reversed b-cell dedifferentiation.
Ever since b-cell dedifferentiation was recognized as an important
contributing factor in b-cell dysfunction in T2DM [20], much effort
has been made to induce dedifferentiated b cells back to a mature
and functional state. For example, several studies have reported the
successful reversion of b-cell dedifferentiation by food/calorie
restriction [12,21], in accordance with the clinical evidence that diet
restriction can promote the recovery of islet function in T2DM
patients [7,22]. However, long-term maintenance of T2DM remission
by food/calorie restriction remains a clinical challenge [23,24]. The
current literature suggests that food/calorie restriction may shift
energy balance and hormonal regulation of weight toward weight
gain after weight loss, bringing about deleterious effects on body
518 B. Li et al. / Cytotherapy 23 (2021) 510 520
composition and physiology [25]. Weight loss surgery has proved
effective in ameliorating the extent of b-cell dedifferentiation in
obese [26] and non-obese [27] T2DM rats. However, significant post-
operative complications, such as anastomotic leak or hemorrhage,
dumping syndrome and worsening acid reflux, have limited the clini-
cal application of weight loss surgery in reviving dedifferentiated b
cells and treating T2DM [28,29]. Insulin therapy has favorable out-
comes on recovery and maintenance of islet function in newly diag-
nosed T2DM patients, as it can alleviate glucotoxicity, reduce the
excessive demand on damaged b cells and provide a rest for b cells
[6]. However, the reversal effect of insulin on b-cell dedifferentiation
is controversial. Wang et al. [30] found that insulin therapy promoted
the redifferentiation of dedifferentiated Ngn3+/insulin b cells into
normal and functional Ngn3 /insulin+ b cells in diabetic adenosine
triphosphate-sensitive potassium channel gain-of-function mice,
whereas Ishida et al. [12] found that although insulin treatment low-
ered blood glucose, it had little effect on b-cell dedifferentiation in
16-week-old db/db mice. These conflicting results suggest that
relieving glucotoxicity is insufficient to reverse b-cell dedifferentia-
tion, at least in db/db mice.
UC-MSC therapy has led to great achievements in treating T2DM.
Recently, Wang et al. [15] found that UC-MSCs improved b-cell dys-
function and reversed b-cell dedifferentiation in 7- to 9-week-old
db/db mice and proposed MSC administration as a potential approach
to reversing b-cell dedifferentiation. In the present study, the authors
demonstrated that UC-MSC therapy reversed b-cell dedifferentiation
and improved islet function in HFD/STZ-induced T2DM mice, 12-
week-old db/db mice and 30-week-old db/db mice, enriching the
experimental evidence that UC-MSCs can reverse b-cell dedifferenti-
ation and highlighting the potential of UC-MSCs in ameliorating
T2DM. However, the duration of the therapeutic effect of UC-MSC
treatment requires further clarification in future research.
Clinically, the development of T2DM is a gradual and staged pro-
cess [31]. Likewise, b-cell dedifferentiation is also characterized by
stages [12,32]. In the early stage, prolonged metabolic stress in the
T2DM condition triggers the loss of b-cell-defining transcriptional
factors, key components responsible for glucose-stimulated insulin
secretion and the increase in ALDH1A3 expression. In the late stage,
b cells regress to endocrine progenitor-like states and Ngn3+ cells
emerge in the islets. Db/db mice are a widely used human T2DM
model for studying the mechanism of b-cell dedifferentiation [33,34]
and exploring strategies to redifferentiate b cells in T2DM
[21,35 37]. However, the db/db mice used in these studies were
10 20 weeks old and in the early stage of b-cell dedifferentiation.
For example, Sheng et al. [21] showed in 12-week-old db/db mice
that 3-month calorie restriction maintained chronic euglycemia and
significantly reduced b-cell dedifferentiation. The expression of
Nkx6.1 and Pdx1 in b cells significantly increased and returned to
nearly normal levels in the study. Ishida et al. [12] compared in 16-
week-old db/db mice the efficacy of 1-month dietary restriction,
insulin, phloridzin or rosiglitazone in reducing blood glucose levels
and reversing b-cell dedifferentiation. The results showed that
although all treatments were equally efficacious in ameliorating
hyperglycemia, only diet restriction resulted in a reversion of b-cell
dedifferentiation, manifested by an increase in insulin+ b cells (from
56% to 68%), a 17% increase in FOXO1+ b cells and a decrease in
ALDH1A3+ b cells (from 80% to 25%).
In the present study, the authors examined the reversal effect of
UC-MSCs on early-stage b-cell dedifferentiation in 12-week-old db/
db mice and innovatively explored the reversibility of late-stage
b-cell dedifferentiation by UC-MSCs in 30-week-old db/db mice. In
the 12-week-old db/db mice, UC-MSC infusions markedly alleviated
b-cell dysfunction and significantly reversed b-cell dedifferentia-
tion, manifested by remarkably increased proportions of insulin+ b
cells (from 26.9 § 2.1% to 49.1 § 3.3%), increased Pdx1+/insulin+ cells
(from 29.3 § 1.8% to 41.7 § 2.9%) and decreased ALDH1A3+ cells
(from 30.3 § 4.5% to 7.1 § 1.6%). Compared with the 1-month diet
restriction in the study by Ishida et al. [11], six UC-MSC infusions in
the authors’ current study produced similar efficacy in reducing
ALDH1A3 expression but had greater efficacy in increasing insulin
expression in b cells. Compared with the 3-month calorie restriction
in the study by Sheng et al. [21], which increased the expression of
key transcriptional factors to nearly normal levels, six UC-MSC infu-
sions in the authors’ current study did not increase the expression
of these transcriptional factors to normal levels. In 30-week-old db/
db mice, six UC-MSC infusions mildly attenuated b-cell dysfunction
and partially reversed b-cell dedifferentiation, manifested by
markedly reduced Ngn3+ cells, significantly increased Pdx1 expres-
sion and slightly decreased ALDH1A3 expression in the islets. Com-
pared with the early treatment group, the late treatment group
showed less reversion of b-cell dedifferentiation. This may be partly
explained by the fact that the dedifferentiated b cells in the late
treatment group had reverted to endocrine progenitor cells and
were more primitive than the b cells in the early treatment group.
Increasing the dose of UC-MSCs or the infusion times in the late
treatment group may help to promote redifferentiation. However, it
is possible that there may be a stage in the process of dedifferentia-
tion after which the dedifferentiated b cells are no longer reversible.
This may contribute to the lower reversibility of late-stage b-cell
dedifferentiation. Regardless, the partial reversion of late-stage
b-cell dedifferentiation by UC-MSCs suggests that UC-MSC therapy
may become an alternative treatment for patients with chronic
hyperglycemia.
The major role of glucotoxicity in b-cell dedifferentiation has been
well established [32,38,39]. Wang et al. [30] demonstrated that resto-
ration of normoglycemia and alleviation of glucotoxicity by insulin
therapy were enough to reverse b-cell dedifferentiation in diabetic
adenosine triphosphate-sensitive potassium channel gain-of-func-
tion mice. In the current study, the reversal effect of UC-MSCs on
early- and late-stage dedifferentiation in db/db mice showed that (i)
the improvement in glucose homeostasis was better in the early
treatment group than the late treatment group, (ii) the reduction in
lipid levels was similar between the two groups and (iii) the restora-
tion of b-cell identity was better in the early treatment group than
the late treatment group. The authors’ results appeared to support
the opinion that alleviation of glucotoxicity was the decisive factor in
reversing b-cell dedifferentiation. However, results from Ishida et al.
[12] showed that in db/db mice, although insulin and phloridzin
treatment lowered blood glucose, neither treatment could reverse
b-cell dedifferentiation, suggesting that mitigating glucotoxicity did
not suffice to reverse b-cell dedifferentiation in db/db mice. In com-
parison, diet restriction attenuated hyperglycemia and reversed
b-cell dedifferentiation at the same time. Since diet restriction
exerted an improved effect on blood lipid profiles compared with
insulin or phloridzin, it can be inferred that diet restriction possibly
acted by alleviating glucotoxicity and lipotoxicity so as to relieve
metabolic inflexibility and facilitate reversion of b-cell dedifferentia-
tion. Based on the aforementioned analysis, the authors postulated in
the current study that the simultaneous alleviation of glucose and
lipid disorders in db/db mice by UC-MSC infusions contributed to the
reversal of b-cell dedifferentiation. The unique capacity of UC-MSC
therapy to mitigate glucolipotoxicity makes it a promising strategy
for reviving dedifferentiated b cells and rejuvenating their function-
ality in T2DM treatment.
Additionally, recent in vitro studies using human pancreatic islets
have discovered that blocking key mediators of the Notch [40], trans-
forming growth factor beta [37,41] and Wnt [42] pathways rescues b
cells from dedifferentiation. Since UC-MSCs possess the ability to
secrete a variety of cytokines and growth factors in the tissue repair
process [43], it is possible that certain cytokines or growth factors
may act on one of the aforementioned pathways and redifferentiate
b cells in T2DM, exerting an independent reversal effect on b-cell
 B. Li et al. / Cytotherapy 23 (2021) 510 520
dedifferentiation beyond the improvement in glucose and lipid
homeostasis. Further research is needed to investigate the indepen-
dent role of cytokines or growth factors secreted by UC-MSCs in
reversing b-cell dedifferentiation in T2DM.
Conclusions
By tracking the process of b-cell dedifferentiation and performing
UC-MSC infusions in type 2 diabetic mice, the authors found that UC-
MSCs demonstrated remarkable effects in the reversal of b-cell dedif-
ferentiation, and the reversal effect was better in early- compared
with late-stage b-cell dedifferentiation. In other words, late-stage
b-cell dedifferentiation could still be partially reversed. In db/db
mice with concomitant T2DM and hyperlipidemia, the reversion of
b-cell dedifferentiation “benefitted” from the simultaneous allevia-
tion of glucose and lipid metabolism disorders mediated by UC-
MSCs. The authors’ findings highlight the potential of UC-MSCs in
rescuing dysfunctional b cells in T2DM through the reversal of b-cell
dedifferentiation. Further studies are necessary to investigate the
molecular mechanism of UC-MSC therapy in reversing b-cell dedif-
ferentiation.
Funding
This study was supported by the National Basic Science and
Development Program (81700679, 81700680, 81870578 and
81900704).
Declaration of Competing Interest
The authors have no commercial, proprietary or financial interest
in the products or companies described in this article.
Author Contributions
Conception and design of the study: BL, YC, LZ and YM. Acquisi-
tion of data: BL, YC, YY and JX. Analysis and interpretation of data: SY,
JG and JL. Drafting or revising the manuscript: BL, YC, LZ and YM. All
authors have approved the final article.
Acknowledgments
This study was supported by the National Basic Science and
Development Program (81700679, 81700680, 81870578 and
81900704).The authors thank Zhiqiang Wu and Qian Mei for provid-
ing technical assistance and Junyan Zou, Zhengyuan Gong, Kun Zhao
and Linxi Zhang for insightful discussions regarding this work.
Supplementary materials
Supplementary material associated with this article can be found
in the online version at doi: 10.1016/j.jcyt.2021.01.005.
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