DERIVATION OF PLURIPOTENTIAL EMBRYONIC STEM-CELLS FROM MURINE PRIMORDIAL GERM-CELLS IN CULTURE
DERIVATION OF PLURIPOTENTIAL EMBRYONIC STEM-CELLS FROM MURINE PRIMORDIAL GERM-CELLS IN CULTURE
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DOI：
10.1016/0092-8674(92)90317-6
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发表时间：
1992-09-04
期刊：
影响因子：
64.5
通讯作者：
HOGAN, BLM
中科院分区：
文献类型：
--
作者：
MATSUI, Y;ZSEBO, K;HOGAN, BLM
钢因子（SF）和白血病抑制因子（LIF）协同促进小鼠原始生殖细胞（PGCs）的增殖和存活，但仅在培养的有限时间内。我们在这里表明，在存在膜相关SF和LIF的情况下，向培养物中加入bFGF可增强PGCs的生长，并使其在正常停止体内分裂的时间之后继续增殖。它们形成密集堆积的、碱性磷酸酶阳性、SSEA-1阳性细胞的集落，在形态上类似于未分化的胚胎干（ES）细胞。这些培养物可以在饲养层上维持至少20代，并且在适当的条件下在单层培养物和裸鼠的肿瘤中产生胚状体和多种分化的细胞表型。PGC衍生的ES细胞在注射到宿主囊胚中时也可以有助于嵌合体。PGCs的长期培养及其向多能ES细胞的重编程对生殖细胞生物学和畸胎瘤的诱导具有重要意义。
Steel factor (SF) and LIF (leukemia inhibitory factor) synergistically promote the proliferation and survival of mouse primordial germ cells (PGCs), but only for a limited time period in culture. We show here that addition of bFGF to cultures in the presence of membrane-associated SF and LIF enhances the growth of PGCs and allows their continued proliferation beyond the time when they normally stop dividing in vivo. They form colonies of densely packed, alkaline phosphatase-positive, SSEA-1-positive cells resembling undifferentiated embryonic stem (ES) cells in morphology. These cultures can be maintained on feeder layers for at least 20 passages, and under appropriate conditions give rise to embryoid bodies and to multiple differentiated cell phenotypes in monolayer culture and in tumors in nude mice. PGC-derived ES cells can also contribute to chimeras when injected into host blasto-cysts. The long-term culture of PGCs and their reprogramming to pluripotential ES cells has important implications for germ cell biology and the induction of teratocarcinomas.