Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells
Derivation of human embryonic stem cell lines in serum replacement medium using postnatal human fibroblasts as feeder cells
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DOI：
10.1634/stemcells.2004-0201
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发表时间：
2005-04-01
期刊：
影响因子：
5.2
通讯作者：
Hovatta, O
中科院分区：
文献类型：
--
作者：
Inzunza, J;Gertow, K;Hovatta, O
没有动物源性材料的人胚胎干细胞（hESC）的衍生和培养将是细胞移植的最佳选择。我们用血清替代（SR）培养基代替传统的胎牛血清和人包皮成纤维细胞作为饲养细胞，获得了两个新的hES（HS 293和HS 306）和10个早期细胞系。自2003年10月以来，HS 293系一直处于连续培养中，传代时间为5-8天，传代水平为56。自2004年2月以来一直培养细胞系HS 306，目前为第41代。所述细胞系表达多能hESC的标志物（Oct-4、SSEA-4、TRA-1-60、TRA-1-81、GCTM-2和碱性磷酸酶）。多能性已在体外拟胚体中显示，并且通过严重联合免疫缺陷/米色小鼠中的畸胎瘤形成也在体内显示了品系293的多能性。HS 293的核型为46，XY，HS 306的核型为46，XX。自2004年9月以来，在类似条件下又获得了10条早期品系。我们的结论是，人胚胎干细胞系可以成功地使用SR培养基和出生后的人成纤维细胞作为饲养细胞。这是迈向无异种条件的一步，并促进了这些细胞在移植中的使用。
Derivation and culture of human embryonic stem cells (hESCs) without animal-derived material would be optimal for cell transplantation. We derived two new hES (HS293 and HS306) and 10 early cell lines using serum replacement (SR) medium instead of conventional fetal calf serum and human foreskin fibroblasts as feeder cells. Line HS293 has been in continuous culture, with a passage time of 5-8 days, since October 2003 and is at passage level 56. Line HS306 has been cultured since February 2004, now at passage 41. The lines express markers of pluripotent hESCs (Oct-4, SSEA-4, TRA-1-60, TRA-1-81, GCTM-2, and alkaline phosphatase). The pluripotency has been shown in embryoid bodies in vitro, and the pluripotency of line 293 has also been shown in vivo by teratoma formation in severe combined immuno-deficiency/beige mice. The karyotype of HS293 is 46,XY, and that of HS306 is 46,XX. Ten more early lines have been derived under similar conditions since September 2004. We conclude that hESC lines can be successfully derived using SR medium and postnatal human fibroblasts as feeder cells. This is a step toward xeno-free conditions and facilitates the use of these cells in transplantation.