Derivation and characterization of human embryonic germ cells: serum-free culture and differentiation potential
Derivation and characterization of human embryonic germ cells: serum-free culture and differentiation potential
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DOI：
10.1016/s1472-6483(10)60079-x
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发表时间：
2009-08-01
影响因子：
4
通讯作者：
Gao, Zhimin
中科院分区：
文献类型：
--
作者：
Hua, Jinlian;Yu, Haisheng;Gao, Zhimin
本研究考察了化学成分确定的培养基补充剂——敲除血清替代物（KSR）对人胚胎生殖细胞（hEGC）生长和分化的影响，发现在 KSR 培养基中初始建立 hEGC 系的效率显着高于在胎牛血清（FCS）培养基中。在 KSR 培养基中生长的未分化 hEGC 集落的百分比显着高于基于 FCS 的培养基（P < 0.05）。 hEGC 集落显示出典型的小鼠胚胎生殖细胞样形态。它们表现出正常且稳定的二倍体核型，并表达碱性磷酸酶（AP）、阶段特异性胚胎抗原（SSEA）和其他多能细胞的特异性标志物。此外，hEGC可以形成简单的囊状胚状体（EB），其由各种细胞类型组成，包括神经细胞、上皮细胞和有节律跳动的心肌细胞，甚至精子样细胞和卵母细胞样细胞。在裸鼠体内形成肿瘤样生长物，并发现其含有多种细胞类型，包括子宫上皮、脂肪细胞、鳞状组织和皮肤结构。总之，我们已经开发出适合 hEGC 系建立的无血清培养系统。这可能提供一个体外模型来研究分化，并可用作不孕症和再生医学治疗的潜在来源。
This study examined the effects of a chemically defined culture medium supplement, knock-out serum replacement (KSR), on the growth and differentiation of human embryonic germ cells (hEGC) and found that the efficiency of the initial establishment of hEGC lines in KSR medium was significantly higher than in fetal calf serum (FCS) medium. The percentage of undifferentiated hEGC colonies growing in KSR medium was significantly higher than in FCS-based medium (P < 0.05). The hEGC colonies showed typical mouse embryonic germ cell-like morphology. They showed normal and stable diploid karyotype and expressed alkaline phosphatase (AP), stage-specific embryonic antigens (SSEA) and other specific markers of pluripotent cells. In addition, hEGC could form simple and cystic embryoid bodies (EB) that consisted of various cell types including neural, epithelial and rhythmically beating cardiac cells, even sperm-like and oocyte-like cells. Tumour-like outgrowths were formed in nude mice and found to contain a variety of cell types, including uterine epithelium, adipocytes, squamous tissue and skin structures. In conclusion, an appropriate serum-free culture system has been developed for the establishment of hEGC lines. This may provide an in-vitro model to study differentiation and can be used as a potential source of therapy for infertility and regenerative medicine.