Abstract
The human embryonic stem cell line RCe010-A (RC-6) was derived from a frozen and thawed blastocyst voluntarily donated as unsuitable and surplus to fertility requirements following ethics committee approved informed consent under licence from the UK Human Fertilisation and Embryology Authority. The cell line shows normal pluripotency marker expression and differentiation to the three germ layers in vitro. It has a normal 46XY male karyotype and microsatellite PCR identity, HLA and blood group typing data are available.
Highlights
RCe010-A (RC-6) was derived from a frozen and thawed blastocyst that was surplus to requirement or unsuitable for clinical use
A microsatellite PCR profile has been obtained for the cell line, and Human Leukocyte Antigen (HLA) Class I and II typing is available (Table 2)
Derivation of hESC from surplus to requirement and failed to fertilise/ develop embryos was approved by The Scotland A Research Ethics Committee and local ethics board at participating fertility clinics and conducted under licence no R0136 from the UK HFEA with informed donor consent
Summary
RCe010-A (RC-6) was derived from a frozen and thawed blastocyst that was surplus to requirement or unsuitable for clinical use. RCe010-A (RC-6) was shown to be pluripotent by expression of Oct-4, Nanog, Tra-1-60 and SSEA-4, but low levels of SSEA-1, using immunocytochemistry (Table 1, Fig. 1). The expression of pluripotency makers Oct 4 and SSEA-4 was 86.2% and 96.8%, respectively, whereas low expression of the differentiation marker SSEA-1 (5.9%) was observed (Fig. 2). Differentiation to the three germ layers, endoderm, ectoderm and mesoderm, was demonstrated using embryoid body formation and expression of the germ layer markers α-fetoprotein, β-tubulin and muscle actin (Fig. 3). A microsatellite PCR profile has been obtained for the cell line, and HLA Class I and II typing is available (Table 2). DNA Profiling to give cell line its signature, gender/species To assess levels of staining for the pluripotency markers Assess antigen levels & cell surface markers commonly associated with hESC
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