Celltissue Bank
We developed the 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 morulas, the zona pellucida is removed and the morula is 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 became possible with the introduction of PGD: the mutation-free embryos are transferred to the woman's uterus, while those that are affected provide a valuable source of ES cell lines with genetic abnormalities. 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 Available 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 inquire online. Also visit StemRide International.