[Culture of porcine mesenchymal stem cells in vitro and developmental capacity of reconstructed embryos by nuclear transfer].
The developmental potential of reconstructed embryos varied according to the source of donor cells, it was thought that the donor cells capabilities to be reprogrammed were different. We established the method of culturing porcine bone marrow mesenchymal stem cells (pMSCs), identified and observed the growth characteristics of pMSCs, and determined pMSCs reprogramming potential as donor cells for nuclear transfer (SCNT). We found that the method of gradient centrifugation to isolate pMSCs from porcine bone marrow was better than the method of anchoring culture; the number of pMSCs achieved peak at day 6, the adhesive rate of cultured cells was 78.50% at 10h and the division index of cultured cells was 24.00 per thousand at day 4. The developmental competence were compared among three kinds of embryos, reconstructed embryos with PF and pMSCs, Parthenogenetic. The blastocysts rate and total cell number of blastocysts were 15.07%, 14.63% vs 30.91% and 24.1 +/- 6.5, 30.67 +/- 17.7 vs 25.8 +/- 11.4. These results indicated that pMSCs could be high proliferation and stable growth characters in vitro, and were suitable donor cells type for nuclear transfer.
Wang
Zhen Kun
ZK
College of Life Science, Northeast Agricultural University, Harbin 150030, China.
Tian
Jiang Tian
JT
Li
Qiu Ming
QM
Wu
Mei Ling
ML
Kong
Qing Ran
QR
Zheng
Zhong
Z
Yu
Xue Wu
XW
Lei
Lei
L
Liu
Zhong Hua
ZH
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D046149:Embryo Culture Techniques / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology*; D053652:Nuclear Transfer Techniques; D013552:Swine