Effects of gestational age on proliferative and differentation potency of mesenchymal stem cells isolated from canine amnion and umbilical cord matrix

ABSTRACT - Amniotic membrane (AM) and umbilical cord matrix (UCM) mesenchymal stem cells (MSCs) have been isolated and characterized in humans and large animal models. In order to distinguish which cells retain the best features for different purposes, the effects of gestational age on proliferation and differentiation potency of canine AM-MSCs and UCM-MSCs was analyzed. Samples were recovered after elective ovariohysterectomy from bitches in early (35 to 40 days) and late (45 to 55 days) fetal stage of pregnancy. The proliferation study and the molecular analysis of embryonic, mesenchymal and hematopoietic markers were performed. Cell neurogenic and osteogenic differentiation were followed. No differences were noticed when comparing data obtained from cells isolated at different gestational ages. Doubling times, cell viability and Oct-4, CD29 and CD44 stemness markers expression were similar in cell isolated from bitches in early or late pregnancy. In both gestational ages, morphological features of neuronal and osteogenic differentiation were observed which need to be confirmed by molecular analysis. In conclusion, our data indicate the possibility to isolate MSCs from canine fetuses at early and late gestational ages with the same proliferative and differentiative capabilities.