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In Search of Common Ground: Search and You Will Find
In Search of Common Ground: Search and You Will Find
The explosion in stem-cell research that fol- established in 2005 to characterize, hold and stem-cells lines derived at the company. Cel-
lowed the isolation of human embryonic stem distribute these approved human embryonic lartis in Gothenburg, Sweden, and Millipore
cells1 in 1998 has seen the number of cell lines stem-cell lines. For each of these lines, the bank in Billerica, Massachusetts, also provide cell
available to researchers increase dramatically. is currently performing extensive testing, says lines. Cellartis is one of the largest sources for
This burst may be due to the fact that embry- Hei, including characterization of gene expres- defined embryonic stem-cell lines, boasting 30
onic stem cells are pluripotent having the sion profiles, karyotype stability, and other different lines that can be obtained at various
potential to generate all adult and embryonic standard embryonic stem-cell assays such as stages of passage or as subclones of selected
cell types so there are excep- lines. Millipore now offers two
INVITROGEN
Engineered stem cells differentiated into 3 tubulin-expressing neurons (red) while continuing to express green fluorescent protein.
culture. Cell lines vary in how easy they are to cells themselves, says Alain Fairbank, research cell lines, Invitrogen is also actively researching
propagate in vitro, says Pera. And Lanza notes market manager at Thermo Fisher Scientific. engineered stem cells. We have been doing a
that researchers at Advanced Cell Technology The company now offers four mesenchymal lot of research to convert stem cells themselves
can tell embryonic stem-cell lines apart based (multipotent stromal cells) stem-cell lines, a into tools, says Goswami. He says that these
on the behaviour of the cells in culture. haematopoietic (germinal cells from umbili- engineered cell lines act as single-cell reporters
cal cord blood) stem-cell line, and a recently providing a visual readout from a live stem cell
The mature way to look at stem cells discovered multipotent cord-blood cell line that as it differentiates to separate lineages.
Although embryonic stem cells are pluripotent, Fairbank says seems to be less restricted in its But even as certain adult stem cells become
adult stem cells are multipotent, and can only multi-lineage potential. The advantage of this more readily available with well-validated
regenerate specific adult cell types in the body. approach is that all media, reagents and culture- approaches to culturing and differentiation,
And for those researchers interested in investi- ware are certified and validated to work with there is still much work to be done. Many
gating the potential of adult somatic stem cells, these specific cell lines. Our focus has been adult stem-cell populations remain difficult
several companies are advancing the idea of developing tools that are validated to work to propagate and expand ex vivo, says Pera.
providing adult stem cells that have been quali- together with stem cells, Fairbank says. And with the exception of mesenchymal
fied for culturing on specific media. Invitrogen of Carlsbad, California, is also stem cells, he says, this has not changed dra-
Thermo Fisher Scientific supplied media and heavily focused on mesenchymal research, matically in recent years. Although Goswami
reagents for culturing adult stem cells, but has says Joydeep Goswami, the companys vice- agrees, he also points to another potential
now moved into supplying adult stem-cells lines president of stem-cell research. These cells issue. It is not that adult stem cells have not
as well. Our strategy has not only been to create are probably going to be the first non-hae- been isolated, the bigger issue is how do you
kits to support culturing and differentiation of matopoietic stem cells to be used for treating get well characterized stem cells? He notes
adult stem cells, but also offer the validated stem patients. In addition to providing adult stem- that in some instances researchers will call
INVITROGEN
signals to differentiate to other cell another. The results showed that in a controlled manner, says Mark
lineages. Although certain growth by simply varying the elasticity Powers, a director at Invitrogen.
factors are known to promote of the matrix, the attached Powers says that when embryonic
some differentiation programmes, mesenchymal stem cells could stem cells form embryoid bodies,
it is now becoming clear that undergo either neurogenesis, they can aggregate into very large
physical interactions between cells myogeneis or osteogenesis. They structures where the cells on the
and mechanical sensing may also went on to show that once the cells inside can be oxygen or nutrient
help to promote differentiation. adhere, they begin setting up the limited. But with Algimatrix the
So researchers are developing stress fibres that actively pull on Three-dimensional matrices can aggregates grow to a consistent
a variety of three-dimensional the adhesions and on the matrix promote cell differentiation. size and not beyond the size of the
(3D) matrices for stem-cell outside. We showed that the cells pores provided by the scaffold.
differentiation. feel the matrix and respond to it, grow their cells on. This is And since it is an inert scaffold, it
In 2006, Dennis Discher of the says Discher. important because researchers actually promotes cell interactions.
University of Pennsylvania in Stefan Przyborski is the founder know how cells respond to this A scaffold such as Algimatrix
Philadelphia, and his colleagues, of Reinnervate in Durham, UK, material in two-dimensional would allow a researcher to culture
demonstrated the potential of the developer of a new scaffold applications. But he also notes that cells in 3D aggregates to promote
the matrix alone to promote for routine 3D cell culture. We this 3D environment enhances differentiation.
differentiation. The idea was to create ways of making the in vitro differentiation when compared It is becoming clear that
use synthetic gels with a collagen environment a more realistic with cells cultured on two- mechanical interaction has a role in
monolayer to mimic the elasticities environment for cell growth, he dimensional plates. cell differentiation. What we are
of certain tissues and see how the says. Reinnervates scaffold is Invitrogen of Carlsbad, California really describing is a sense of touch
cells respond, says Discher. The unique, but might seem familiar has developed Algimatrix, an inert the cells have no eyes or ears,
work was done under constant to biologists who have performed 3D scaffold made of alginate. so they use this sense of touch to
serum conditions, without any cell culture. We created a scaffold The idea behind Algimatrix is tell where they are this is all part
discriminating soluble factors or made of polystyrene, the same that it provides a framework for of sensing and responding to the
growth factors that might promote material that people currently cells to reside in that will enable environment, says Discher. N.B.
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NATURE|Vol 451|14 February 2008 TECHNOLOGY FEATURE STEM CELLS
cells a particular type of stem cell, but the next where cellcell interaction can promote Differentiation of embryonic-stem cells or
batch will have different marker profiles, indi- differentiation, Matrigel is coated as a two- adult stem cells into particular lineages is an
cating a different cell type. dimensional scaffold and is therefore effective area of intense research for developmental
for long-term embryonic stem-cell culture. biologists and researchers interested in using
Mouseless developments Although Matrigel is a stem calls for therapeutic
V. CHRISTIE
The culturing of embryonic stem cells has solubilized preparation applications.
often relied on the use of mouse embryonic extracted from a mouse This is where the bat-
fibroblast cells (MEFs) as feeder cells in dif- sarcoma potentially tlefront is, says Lanza,
ferent media formulations containing serum. raising similar issues We need to learn how
It is thought that the MEFs secrete factors that to the use of serum to generate different
promote the growth of embryonic stem cells Kosovsky says that they cell types. He points
in culture. But growing embryonic stem cells are currently working on to haematopoetic stem
using MEFs in undefined media and serum more defined formula- cells, which give rise to
that potentially contains animal products tions of the scaffold. all blood cell types, as an
could limit the potential of stem-cell lines for Invitrogen is one of example of how difficult
therapeutic applications. several companies mov- the task can be. In an
It all comes down to how the regulators ing away from culture embryo these cells start
look at these lines and how they have been conditions that rely on out in the yolk sac, then
handled over the years, says Bruce Davidson, either feeder-cells or travel to the aortic arch,
chief scientific officer at ESI. It is for this rea- serum. We are provid- Reinnervate in Durham, UK, derives to the liver and then to
son that ESI worked to derive clinical-grade ing stem-cell media neurons from human pluripotent stem the bone marrow, and
human embryonic stem cells. We have four with a twist it is a cells using synthetic compounds. at each point along that
GMP- [good manufacturing practice] certified serum-free and defined journey are being edu-
embryonic stem-cell lines that were developed media, says Goswami. This medium, called cated by their surroundings. Replicating this
over the past couple of years and are now ready StemPro hESC SFM, works with 16 embry- is not easy. Advanced Cell Technology has been
for distribution, says Davidson. Cellartis also onic stem-cell lines from around the world. working on the differentiation potential of the
provides a human embryonic stem-cell line Millipore also offers a serum-free medium haemangioblast a precursor to hematopoi-
that was derived without any contact with ani- called HEScGRO, which works with a variety etic cells that they isolated. People have got
mal products. And many more researchers and of embryonic stem-cell lines. other haemangioblast-like cell types, but these
companies are trying to identify the important are KDR-negative and CD31-negative, so they
factors for culturing embryonic stem cells Different paths are an earlier progenitor cell than whats been
and develop robust feeder-free approaches The field is moving towards defined media described previously, which is why they expand
in well-defined media. without animal components and without feed- and do things so much better, says Lanza. Dif-
The WiCell Research Institute in Madison, ers, says Pera. But some scientists are being ferentiating these haemangioblasts has allowed
Wisconsin, has a major focus on optimizing cautious about these culture systems. Although the companys researchers to generate entire
embryonic stem-cell culture media and condi- signs are encouraging, Pera says that the jury is tubes of red blood cells, as well as other hemat-
tions to allow long-term culture of stem cells that still out on most of these new systems. And the opoietic lineages and endothelial cells at very
do not differentiate. In recent years, research- NSCB is taking a careful approach to feeder- high efficiencies.
ers at WiCell have found that small changes to free cultures. We made the decision not to use Although success has also been reported in
either media or the physiochemical conditions some of the feeder-free methods that are com- using differentiation protocols for deriving
of the culture can have a profound effect on the ing out right now, but to stick at this stage to the neural progenitor cells, cardiomyocytes and
viability of cells. But in 2006, WiCell researchers standard MEF-based methods, says Hei. The retinal pigment epithelium from embryonic
reported culturing embry- decision was made in part stem cells, in most cases studies have shown
R. CARNACHAN
onic stem cells without to avoid forcing the research that only a very small percentage of cells dif-
using MEFs, instead relying community towards feeder- ferentiate into a particular tissue type. Some
on protein components that free methods while they are researchers now think that multiple cells or
were either recombinantly still in the early stages of interactions between cells and surfaces might
derived or purified from development. But Hei also be required to obtain a desired derivative from
human materials3. acknowledges that at a later stem cells (see Beyond the flat world).
That medium mTeSR-1 date they might develop But Pera sees this as one area that is stead-
is now marketed by banks of cells using feeder- ily progressing. I think we are getting more
StemCell Technologies in free systems. Although and better differentiation protocols that are
Vancouver, Canada, who some express concerns that more defined and have better endpoints, he
is collaborating with BD cells cultured in feeder-free says. However, he does think that there is lack
Biosciences in San Jose, systems do not thrive as of comparative data on different cell lines and
California, and WiCell well, Goswami is confident their abilities to differentiate using the current
on feeder-free cell culture Poylstyrene scaffolds can support a in these new systems and protocols although the available data would
environments for human variety of cells in culture. says that the time has come indicate that there will be differences between
embryonic stem-cell cul- for feeder-free, serum-free certain cell lines in the ability to perform in
ture using BD Biosciences Matrigel scaffold. embryonic stem-cell culture. Using our media specific differentiation protocols.
Colonies formed on Matrigel hESC-qualified we can get equivalent or higher numbers of
matrix with mTeSR-1 media tend to be spread cells compared with any feeder-dependent Looking up
out and form a monolayer-like morphology, system, he says. The world might soon be slightly easier for
which makes passage easier and transfec- We are still learning a lot about what it researchers working on mesenchymal stem-
tion more efficient, says Marshall Kosovsky, takes to coax a stem cell to become a particu- cell differentiation. Both Thermo Fisher Sci-
technical support manager at BD Biosciences. lar tissue, says George Daley from Harvard entific and Invitrogen are introducing kits in
Unlike three-dimensional growth matrices, Medical School in Boston, Massachusetts. the coming months for the differentiation of
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TECHNOLOGY FEATURE STEM CELLS NATURE|Vol 451|14 February 2008
mesenchymal stem cells to adipogenic (fat- immune-response issues in patients. cells thrived in culture. It turns out that
producing), osteogenic (bone-producing) and As companies move their products closer although you can derive cells on human feeders
chondrogenic (cartilage-producing) lineages. to clinical trials, the questions on everyones or even feeder-free with extracellular matrices,
These kits provide all the necessary growth mind are when and under what conditions they do not thrive as well, says Lanza. And for
factors to direct the differentiation of the mes- will the US Food and this reason he thinks
858