Isolation, characterization, and gene modification of dairy goat mesenchymal stem cells from bone marrow

Abstract

Bone marrow mesenchymal stem cells (MSCs) are adult pluripotent cells that are considered to be an attractive cell type for therapy models and for nuclear transfer transgenesis. To date, MSCs from various species have been studied, but only a limited amount of information regarding dairy goat MSCs (gMSCs) is available. The objectives of this study were to isolate, induce the multilineage mesenchymal differentiation, and investigate the gene modification efficiency of gMSCs, thereby initiating further research on these cells. The gMSCs isolated from bone marrow grew, attached to plastic with a typical fibroblast-like morphology, and expressed the mesenchymal surface marker CD44, CD29, CD90, and CD166, but not the hematopoietic marker CD45. Furthermore, the gMSCs expressed the transcription factors Oct-4 and Nanog, which have been shown to be critical for stem cell self-renewal and pluripotency. The multilineage differentiation potential of gMSCs was revealed by their ability to undergo adipogenic and osteogenic differentiation when exposed to specific inducing conditions. Transient transduction of gMSCs with a plasmid containing the GFP gene resulted in higher transfection rate compared with fetal fibroblasts (FFs). Furthermore, cell colonies with stable genetic modifications were obtained when gMSCs were transfected with a mammary-specific expression vector containing human lysosomal acid beta-glucosidase gene (hGCase). In conclusion, these results demonstrated that typical mesenchymal stem cells were isolated from dairy goat bone marrow, possessed the characteristics of pluripotent stem cells, and had the potential of specific genetic modifications for gene therapy and producing transgenic goats.