Hofmann M C, Hess R A, Goldberg E, Millán J L
La Jolla Cancer Research Foundation, Cancer Research Center, CA 92037.
Proc Natl Acad Sci U S A. 1994 Jun 7;91(12):5533-7. doi: 10.1073/pnas.91.12.5533.
Establishing mammalian germ-cell lines capable of differentiation in vitro would greatly facilitate the study of gametogenesis and the meiotic process that is so fundamental for reproduction and the maintenance of genetic diversity of the species. We have established two germ-cell lines [GC-2spd(ts) and GC-3spc(ts)] by cotransfecting primary mouse testicular germ cells with the simian virus 40 large tumor antigen gene and the gene coding for a temperature-sensitive mutant of p53. Both cell lines express the germ cell-specific lactate dehydrogenase C4 isozyme and cytochrome ct isoform. At the permissive temperature of 37 degrees C, the GC-2spd(ts) line generates cells with a haploid DNA content and morphologic and biochemical features of round spermatids, including the appearance of an acrosomic granule. The identification of a flagellar axoneme when these cells are cultured at 32 degrees C further indicates that these cells correspond to the early spermatid stages of spermiogenesis.
建立能够在体外分化的哺乳动物生殖细胞系将极大地促进配子发生和减数分裂过程的研究，而减数分裂过程对于物种的繁殖和遗传多样性的维持至关重要。我们通过将猿猴病毒40大肿瘤抗原基因和编码p53温度敏感突变体的基因共转染到原代小鼠睾丸生殖细胞中，建立了两个生殖细胞系[GC-2spd(ts)和GC-3spc(ts)]。这两个细胞系均表达生殖细胞特异性乳酸脱氢酶C4同工酶和细胞色素ct异构体。在37℃的允许温度下，GC-2spd(ts)系产生具有单倍体DNA含量以及圆形精子细胞形态和生化特征的细胞，包括顶体颗粒的出现。当这些细胞在32℃培养时鞭毛轴丝的鉴定进一步表明这些细胞对应于精子发生的早期精子细胞阶段。
