Enhanced development of porcine embryos cloned from bone marrow mesenchymal stem cells

Abstract

In the present study, we have characterized an isolated population of porcine bone marrow mesenchymal stem cells (MSCs) for multilineage commitment and compared the developmental potential of cloned embryos with porcine MSCs and fetal fibroblasts (FFs). MSCs exhibited robust alkaline phosphatase activity and later transformed into mineralized nodules following osteoinduction. Furthermore, MSCs underwent adipogenic and chondrogenic differentiation by producing lipid droplets and proteoglycans, respectively. Primary cultures of FFs from a female fetus at similar to 30 day of gestation were established. Donor cells at 3-4 passage were employed for nuclear transfer (INIT). Cell cycle analysis showed that the majority of MSCs in confluence were in the G0/G1 stage. Cumulus-oocyte complexes were matured and fertilized in vitro (IVF) as control. The cleavage rate was significantly (P < 0.05) higher in IVF than in NT embryos with MSCs and FFs (84.5 +/- 4.6% vs. 52.2 +/- 5.4% and 50.8 +/- 5.2%, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs (20.6 +/- 2.5% and 18.4 +/-.0%) did not differ, but were significantly (P < 0.05) higher than NT derived from FFs (9.5 +/- 2.1%). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs (34.4 +/- 5.2 and 0.38 +/- 0.08, respectively) were significantly (P < 0.05) higher than those from FFs (22.6 +/- 5.5 and 0.18 +/- 0.12, respectively). Proportions of TUNEL positive cells in NTembryosfrom FFs (7.3 +/- 1.8%) were significantly (P < 0.05) higher than in MSCs (4.6 +/- 1.3%) and IVF (2.5 +/- 0.9%). The results clearly demonstrate that multipotent bone marrow MSCs have a greater potential as donor cells than FFs in achieving enhanced production of cloned porcine embryos.