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Cell And Tissue Bank
We developed an original technique for the establishment of ES cell lines from human
embryos at the morula stage, which were shown to meet the NIH criteria. To establish
human ES cell lines from morulae, the zona pellucida is removed and the morula placed
under a middle density feeder layer. Within several days, cells outgrow and spread into
the feeder layer. The primary cell disaggregation is performed with EDTA or EGTA, and the
loose cells are transferred back to the feeder layer to proliferate. Fast proliferating
colonies are isolated and propagated further.
No morphological differences between human ES-cells originating from ICM and from morula
were observed, except for being more heterogeneous, as well as the pattern of above marker
expression. The established human ES-cell lines are maintained in vitro from 10 to 15
passages before freezing in sufficient amounts with control thaw out. There are also no
differences in the efficiency of obtaining ES cell lines depending on the source of
preimplantation embryo.
To induce the differentiation of human ES cells in different cell types, the cells are
cultured to form embryonic bodies, which are isolated with subsequent disaggregation and
plating of clusters of cells. The cultured clusters of cells show a wide range of cell
types belonging to ectoderm, endoderm and mesoderm, and spontaneously differentiate in vitro
into a variety of cell types, including neuron-like cells. Controlling differentiation into
pure populations of specific neural cells may eventually form the basis of therapy for some
neurodegenerative disorders and spinal injuries. These developments provide an obvious
potential for the therapeutic use of the embryonic stem cells in clinical practice.
The above developments have made it possible to initiate the establishment of a repository
of ES cell lines with different genetic abnormalities. Although initially the major goal of
the establishment of human ES cell lines was the development of the cell-replacement
therapies, it is presently obvious that human ES cell lines will have an important role in
the studies of mechanisms of genetic disorders through generating the sources of normal and
genetically abnormal cells and tissues.
The establishment of ES cell lines with genetic disorders has become possible with
introduction of PGD, so that the mutation free embryos are transferred while those
affected provide a valuable source of ES cell lines with genetic abnormalities, using the
above techniques for the initiation of ES cell lines from preimplantation embryos. This also
provides a unique opportunity to investigate the potential of establishing ESC lines
depending on the genotype.
Based on our ongoing PGD work described elsewhere (Verlinsky & Kuliev, 2000), we attempted
to establish ES cell lines from embryos with a variety of single gene and chromosomal
disorders. PGD is an important source for the ES cell lines, because the embryos obtained
from PGD are well tested, with the genotype of the potential ES cell line known from the
onset. Our current experience shows that the efficiency of the derivation of ES cell lines
from the embryos with single gene disorders is not affected. This is demonstrated by the
establishment of the first ES cell line repository with different genetic abnormalities,
which presently contains more than 50 human ES cell lines. This repository contains human
ES cell lines obtained from embryos with single gene disorders, such as thalassemia (HBB),
neurofibromatosis type I (NF1), Marfan syndrome (FBN1), myotonic dystrophy (BMD), fragile-X
syndrome (FMR1), and Huntington disease (HD).
For Further Reading
National
Institute of Health Guidelines for Research Using Human Pluriopotent Stem Cells.
NIH Stem Cells Information Archives, July, 2004
Tompson JA, Itskovitz-Eldor J, Shapiro SS, et. al. Embryonic stem cells lines derived
from human blastocysts. Science 1998; 282:1145-1147
What stem cell lines are available?
Below, we have listed the available stem cell lines.
In the Stem Cell Line Avalable column
listed below we provided, where possible, a link to a website with
more information on the genetic disorder, and a link to a medical
website under "Link".
Next Steps:
If you are interested in a RGI's Cell and Tissue Bank your next step
should be to contact us at 773.472.4900 or e-mail
us with any questions of
inquiries. Also visit StemRide International
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