Derivation of goat embryonic stem cell-like cell lines from in vitro produced parthenogenetic blastocysts

Abstract

Parthenogenesis is the biological phenomenon by which embryonic development is initiated without male contribution. Parthenogenesis is a very useful method of derivation of embryonic stem cells (ESCs), which may be an important source of histocompatible cells and tissues for cell therapy. The aim of the present study was to derive and characterize goat embryonic stem cell-like cells from in vitro developed blastocysts following parthenogenetic activation of goat oocytes. Two parthenogenetic embryonic stem cell-like cell (gPESC) lines were established. The gPES cell-like cell colonies showed typical ESC morphology and expressed ESC specific markers such as alkaline phosphatase, Oct-4, Sox-2, Nanog, TRA-1-60, SSEA-4 and TRA-1-81. Genetic stability of the cell lines was proven to be preserved via karyotype analysis which showed normal karyotype. In suspension culture in absence of feeder layer and without LIF, the embryonic stem cell-like cells produced embryoid bodies and following prolonged culture differentiated into several types of cells including neuron like, epithelium like and cardiac muscle cells. Cell specific markers were observed all cells expressed positive marker genes. All of these results demonstrated the feasibility to isolate and establish goat parthenogenetic ES cell-like cell lines, which provides an important tool for studying epigenetic effects in ESCs and developmental biology as well as gene targeting to produce genetically modified livestock.