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Running head: STEM CELL BIOLOGY 1

Stem Cell Biology

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STEM CELL BIOLOGY 2

Guitierrez-Gonsalez et al. (2009) in their work,” Analysis of the clonal architecture of the

human small intestinal epithelium establishes a common stem cell for all lineages and reveals a

mechanism for the fixation and spread of mutations” sought to understand the nature of a

person’s stem cell architecture, Sought to understand the nature of a person’s stem cell, it

architecture, how mutation can be fixed as well as how they are spread. To achieve these

objectives, the researcher considered four main questions. First, does there exists a stem cell

from an adult that can be used in deriving all the differentiated epithelial cells? Secondly, within

a small epithelium, exactly how does a spread in mutation occur? Thirdly, how is the amount of

villi and crypts in a small bowel in a human being maintained? Lastly, as has been observed in

both the stomach and colon, does monoclonal conversion and niche succession happen in the

small intestine crypts. On the other hand, Fellous et al. (2009) in ”Locating the Stem Cell Niche

and Tracing Hepatocyte Lineage in Human Liver” sought to identify hepatocyte deficient

patches in the cytochrome c oxidase enzyme, the component of In both studies, mtDNAs were

utilized as markers for clonal expansion. However, Gutierrez-Gonsalez et al. 2009 in their study

made use of mitochondrial DNA mutation as the marker of stem cell progeny’s clonal expansion.

On the other hand, the mitochondrial DNA mutations were utilized by Fellous et al. 2009 for

clonal expansion of hepatocytes deficient patches.

After obtaining the approval to conduct their study, Guitierrez-Gonsalez et al. used 10

patients with pancreatic tumor in the study. They used normal duodenal specimens particularly

those without dysplasia or showing signs of inflammation. Similarly, Fellous et al. obtained 15

normal fresh frozen human liver blocks after ethical approval to carry out the test.
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For the study by Fellous et al. 2009, immunohistochemistry was carried out on the 4-um

section sliced from the formalin-fixed paraffin bloc of tissue. Immunohistochemistry in

Guiterrez-Gonsalez’s study was conducted on the 2 um paraffin section. They were then

allowed to dry and then dewaxed in xylene. Later they were dried by reducing the alcohol

concentrations. For both Fellous et al. 2009 and Gutierrez-Gonsalez et al.’s study, histochemistry

was done for the Cytochrome C Oxidase Succinate Dehydrogen. The analysis of mtDNA for

both studies was to intend to confirm any clonal expansion.

Guitierrez-Gonsalez et al. 2009 found that the small bowel crypts of human are clonal,

the stem cell is in a position of populating the whole stem cell zone, and that patches of the

crypts have similar mtDNA mutations. Similarly, Fellous et al. found the cytochrome c oxidase-

deficient hepatocytes patches to be clonal implying a root from the stem cell or long-lived cell.

The data from both studies are convincing about the presence of multiple or single cells

in examined region and cell’s clonal expansion in that the hypothesis for both the studies was

confirmed. Moreover, the video on Cre-lox recombination demonstrates through lineage analysis

that it is possible to have these cell in the respective areas of study.

Cre-lox recombination refers to the technology (site-specific recombinase) useful in

conducting inversions, translocations, deletions and insertions in cell DNA. The Cre recombinase

is capable of splicing and recognizing specific DNA sequence referred to as the Loxp Sites. This

LoxP site comprises of thirty-four base pair and 2 bordering palindromic sequences and the eight

bp spacer region.
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References

Gutierrez-Gonsalez, L., Deherogoda, M., Elia, G, Leedham, S., Shankar, A., Imber, C,

Jankowsi, J, Tumbul, D, Novelli, M, Wright, N and McDonald, S. (2009). “Analysis of the

clonal architecture of the human small intestinal epithelium establishes a common stem cell for

all lineages and reveals a mechanism for the fixation and spread of mutations.” J Pathol. 217:

489–496

Fellous, T, Islam, S, Tadrous, P, Elia, G et al. (2009).Locating the Stem Cell Niche and

Tracing Hepatocyte Lineages in Human Liver. Hepatology, Vol. 49, No. X,

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