Generation of Histocompatible Androgenetic Embryonic Stem Cells Using Spermatogenic Cells

Abstract

Androgenetic embryonic stem (aES) cells, produced by pronuclear transplantation, offer an important autologous pluripotent stem cell source. However, the isolation of aES cells, particularly individual-specific aES cells, with the use of fertilized embryos has limited the practical applications of this technology in humans. In this study, we applied a new approach, essentially described as somatic cell nuclear transfer, and generated three aES cell line types with the use of spermatogenic cells including primary spermatocytes, round spermatids, and mature spermatozoa as donor cells, omitting the need to use fertilized embryos. Although abnormality of chimeras and absent germline competency indicated that all three types of aES cells exhibited limited pluripotency, the epigenetic status of the aES cell lines tended to resemble normal ES cells during long-term culture, and some parental-specific imprinted genes were expressed at levels comparable to those of normal ES cells. Furthermore, the histocompatibility of the aES cells was investigated by transplanting the differentiation progenies of the aES cells into major histocompatibility (MHC)-matched and -mismatched recipient mice. The results indicated that these aES cells were histocompatible with MHC-matched mice after transplantation. Our study provides evidence that MHC-competent autologous aES cells could be generated from different spermatogenic cells using nuclear transfer into oocytes, a process that could avoid the use of fertilized embryos. STEM CELLS 2010;28:229–239