DNA fingerprinting confirmed the genetic relation

DNA fingerprinting confirmed the genetic relation of the derivative line to the parental somatic line (Fig. 1B). The PD173074 exhibited normal karyotype (46, XY) upon G-band analysis (Fig. 1C). Pluripotency was verified by gene expression of pluripotent stem cell markers Oct4, Sox2, Nanog, Rex1, DMNT3B and ABCG2 by qPCR (Fig. 1D). In addition, pluripotent surface marker expression at single cell resolution of SSEA3, SSEA4, Tra-1-60, and Tra-1-81 were confirmed by flow cytometry (Fig. 1E). Differentiation capacity into three germ layers was confirmed by PluriTest™ and quantitative PCR (Fig. 2).
Materials and methods
Resource table Resource details We generated KCL018 clinical grade hESC line following protocols, established previously (). The expression of the pluripotency markers was tested after freeze/thaw cycle (). Differentiation potential into three germ layers was verified in vitro (). Materials and methods
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Acknowledgments This work was supported by the UK Medical Research Council grants G0701172 and G0801061. We thank Dr. Yacoub Khalaf, Director of the Assisted Conception Unit of Guy\'s and St Thomas\' NHS Foundation Trust and his staff for supporting the research program. We are especially indebted to Prof Peter Braude and to the patients who donated embryos.
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We generated KCL036 research grade hESC line following protocols established previously (Ilic et al., 2012; Stephenson et al., 2012; Jacquet et al., 2013). The expression of the pluripotency markers was tested after freeze/thaw cycle (Fig. 2; Jacquet et al., 2015). Differentiation potential into three germ layers was verified in vitro (Fig. 3 and Fig. 5; Jacquet et al., 2015) and in vivo (Fig. 4; Jacquet et al., 2015). We also generated research grade of KCL036 line that is adapted to feeder-free conditions (Jacquet et al., 2015).
Materials and methods
Author disclosure statement
Acknowledgments This work was supported by the UK Medical Research Council grants G0701172 and G0801061. We thank Dr. Yacoub Khalaf, Director of the Assisted Conception Unit of Guy\'s and St Thomas\' NHS Foundation Trust and his staff for supporting the research program. We are especially indebted to Prof Peter Braude and to the patients who donated embryos.
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Resource Details RCe009-A (RC-5) was shown to be pluripotent by expression of the pluripotency markers Oct-4, Nanog, SSEA-4, Tra-1-60 and Tra-1-81, but not the differentiation marker SSEA-1, using immunocytochemistry (Table 1, Fig. 1). By flow cytometric analysis, the expression of pluripotency makers Tra-1-60, Tra-1-81 and SSEA-4 was 94.8, 93.6% and 94.8%, respectively, but some expression of the differentiation marker SSEA-1 (37.3%) 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). Blood group genotyping gave the blood group AO1 (Table 2).
Materials and methods
Acknowledgements Research culminating in the derivation of this line was funded by a grant from Scottish Enterprise Economic Development Agency (PM07321, SPO111055) to PDS, MB and AC.
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Verification and authentication
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Verification and authentication
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