Open AccessGrowing of human embryonic stem cells on feeders derived from themselves
Author(s) -
Shi Min An,
Qiao Zeng,
Xiang Teng,
Zi Long,
Juan Li,
Qian Pan,
Ling Wu,
Dong Liang,
Kun Xia,
Jia Xia,
Zhuohua Zhang
Publication year - 2009
Publication title -
yichuan
Language(s) - English
Resource type - Journals
ISSN - 0253-9772
DOI - 10.3724/sp.j.1005.2008.01567
Subject(s) - mesenchymal stem cell , biology , cd90 , embryonic stem cell , homeobox protein nanog , teratoma , embryoid body , stem cell , cd34 , microbiology and biotechnology , pathology , induced pluripotent stem cell , medicine , biochemistry , gene
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.