Introduction

The Stemride International established a repository of more than 150 hESC. The lines were derived by the Reproductive Genetics Institute (RGI). Lines have been grown, using animal and human feeders, from blastocysts and morulae.

RGI developed an original technique generating hESC lines from human embryos at the morula stage, which were shown to meet the NIH criteria. To derive hESC 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 hES cells originating from ICM and from morula were observed, except for being more heterogeneous, as well as the pattern of above marker expression. The hESC 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 hESC lines depending on the source of preimplantation embryo.

The repository contains hESC lines with different genetic and chromosomal abnormalities. These lines will play an important role in studies of genetic disorders mechanisms through generating the sources of normal and genetically abnormal cells and tissues. The hESC lines with genetic disorders have been established by application of the PGD, so that the mutation free embryos are transferred while those affected provide a valuable source of hESC lines with genetic abnormalities. This also provides a unique opportunity to investigate potentials of generating hESC lines depending on the genotype.

PGD is an important source for the hESC lines, because the embryos obtained from PGD are well tested, with the genotype of the potential hESC line known from the onset. The repository contains hESC 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

The bank
All hESC lines in the bank are pluripotent and fully characterized.
The hESC line bank lists all lines and their vital characteristics.

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