Establishment of embryonic stem cell lines from cynomolgus monkey blastocysts produced by IVF or ICSI

Human embryonic stem (ES) cells are predicted to be a valuable source for p roducing ES-derived therapeutic spare tissues to treat diseases by controll ing their growth and differentiation. To understand the regulative mechanis ms of their differentiation in vivo and in vitro, ES cells derived from non human primates could be a powerful tool. We established four ES cell lines from cynomolgus monkey (Macaca fascicularis) blastocysts produced by in vit ro fertilization (IVF) or intracytoplasmic sperm injection (ICSI). The ES c ells were characterized by the expression of specific markers such as alkal ine phosphatase and stage-specific embryonic antigen-4. They were successfu lly maintained in an undifferentiated state and with a normal karyotype eve n after more than 6 months of culture. Pluripotential competence was confir med by the formation of teratomas containing ectoderm-, mesoderm-, and endo derm-derivatives after subcutaneous injection into SCID mice. Differentiati on to a variety of tissues was identified by immunohistochemical analyses u sing tissue-specific antibodies. Therefore, we established pluripotent ES c ell lines derived from monkeys that are widely used as experimental animals . These lines could be a useful resource for preclinical stem cell research , including allogenic transplantation into monkey models of disease. (C) 20 01 Wiley-Liss, Inc.