Transcriptional profiling of rhesus monkey nuclear transfer embryonic stem cells

Derivation of embryonic stem cells (ESC) genetically identical to a patient by somatic cell nuclear transfer (SCNT) holds the potential to cure or alleviate the symptoms of many degenerative diseases while circumventing any immunorejection issues. However, no primate nuclear transfer embryonic stem (ntES) cell lines have been derived to date. Here, we used a modified SCNT technique to produce rhesus macaque SCNT blastocysts at a relatively high efficiency from adult donor cells and we successfully derived two primate ntES cell lines from 304 oocytes (an overall efficiency of 0.7%). Nuclear and mitochondrial DNA analysis confirmed the ntES cell lines were derived from rhesus monkey SCNT blastocysts and both rhesus monkey ntES cell lines exhibited a normal ESC morphology, expressed key stemness markers, were transcriptionally indistinguishable from control ESC lines and differentiated into multiple cell types. This is, to our knowledge, the first confirmed derivation of primate ntES cell lines. Keywords: rhesus monkey somatic cell nuclear transfer embryonic stem cells We compared the transcriptomes of rhesus monkey embryonic stem cell lines derived from IVF-produced embryos (ORMES-10 and ORMES-22) with rhesus monkey nuclear transfer embryonic stem cells derived from somatic cell nuclear transfer produced embryos (CRES-1 and CRES-2). We also analyzed the donor adult somatic skin fibroblast cell line used to produce the cloned embryos. Three biological replicates of each cell line (A, B and C) was analyzed.