Derivation of Human Embryonic Stem Cells from Developing and Arrested Embryos

Abstract

Human embryonic stem cells (hESC) hold huge promise in modern regenerative medicine, drug discovery, and as a model for studying early human development. However, usage of embryos and derivation of hESC for research and potential medical application has resulted in polarized ethical debates since the process involves destruction of viable developing human embryos. Here we describe that not only developing embryos (morulae and blastocysts) of both good and poor quality but also arrested embryos could be used for the derivation of hESC. Analysis of arrested embryos demonstrated that these embryos express pluripotency marker genes such OCT4, NANOG, and REX1. Derived hESC lines also expressed specific pluripotency markers (TRA-1-60, TRA-1-81, SSEA4, alkaline phosphatase, OCT4, NANOG, TERT, and REX1) and differentiated under in vitro and in vivo conditions into derivates of all three germ layers. All of the new lines, including lines derived from late arrested embryos, have normal karyotypes. These results demonstrate that arrested embryos are additional valuable resources to surplus and donated developing embryos and should be used to study early human development or derive pluripotent hESC.

Development of Human Cloned Blastocysts Following Somatic Cell Nuclear Transfer with Adult Fibroblasts

“…The recent findings with mouse [57] and human [58] embryos showing the generation of ESC lines from individual blastomeres in human from arrested embryos [59] and primate embryonic stem cells following somatic cell nuclear transfer [60] may hasten the attainment of this goal.…”

Diploid, but not haploid, human embryonic stem cells can be derived from microsurgically repaired tripronuclear human zygotes

“…The from eight-cell embryos; however, the derivation efficiency was poor. 39,48,49 Here, we showed a much higher efficiency of ES derivation from r2PN embryos, which would be helpful for deriving more human ES cell lines. Moreover, this approach would allow researchers more opportunities to derive human ES cells with certain diseases, such as genetic mutations and metabolic and endocrine diseases.…”

Derivation of human embryonic stem cell lines from single blastomeres of low-quality embryos by direct plating

“…The LQ embryos are discarded. However, this grading method cannot precisely predict the developmental potential of embryos, especially the potential of each blastomere [13,14]. Veiga et al [15] reported that pregnancy can be achieved by replacing a frozen-thawed embryo with less than 50 % intact blastomeres, indicating the full developmental potential of the blastomeres.…”