[Growing of human embryonic stem cells on feeders derived from themselves].
This study was carried out to determine whether mesenchymal stem cells (MSCs) derived from teratoma of human embryonic stem cells (hESCs) function as feeder cells to support hESCs growth. Approximately 5x10(6) hESCs were injected into the hind limb muscle of each SCID-beige mouse to form teratoma. After 8 weeks, the MSCs were isolated from the teratoma and cultured in Mesencult medium. Purified MSCs were then used as the feeder cells for hESCs culture. High purity MSCs derived from teratoma were isolated. The cells were morphologically similar to bone marrow MSCs (bMSCs). The teratoma-derived MSCs were negative for CD34 and CD45 but positive for CD29, CD49b, CD105, CD73, and CD90, which resembled those expressed by bMSCs. After passaged on MSCs feeder cells more than 10 passages, hESCs maintained hESC characteristics in morphology. Reverse PCR showed the expression of Oct4 and Nanog. SSEA-1 was negative and SSEA-4, TRA-1-60, and TRA-1-81 were positive. Alkaline phosphatase staining showed positive results.The karyotype remained normal. Moreover, the hECSs cultured on teratoma-derived MSCs formed teratoma in vivo and embryoid body in vitro confirmed their pluripotency. Accordingly, MSCs derived from hESCs by in vivo differentiation can be used as the feeder cells for hESCs culture.
An
Shi-Min
SM
National Lab of Medical Genetics, Central South University, Changsha 410078, China. csuanshimin@yahoo.com.cn
Zeng
Qiao
Q
Teng
Xiang-Yun
XY
Long
Zhi-Gao
ZG
Li
Juan
J
Pan
Qian
Q
Wu
Ling-Qian
LQ
Liang
De-Sheng
DS
Xia
Kun
K
Xia
Jia-Hui
JH
Zhang
Zhuo-Hua
ZH
D015720:5'-Nucleotidase / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D000071063:Endoglin; D005434:Flow Cytometry; D006801:Humans; D039421:Integrin alpha2 / Q000378:metabolism; D019012:Integrin beta1 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D051379:Mice; D016513:Mice, SCID; D011956:Receptors, Cell Surface / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013724:Teratoma; D018800:Thy-1 Antigens / Q000378:metabolism
D015703:Antigens, CD; C064206:ENG protein, human; D000071063:Endoglin; D039421:Integrin alpha2; D019012:Integrin beta1; D011956:Receptors, Cell Surface; D018800:Thy-1 Antigens; D015720:5'-Nucleotidase