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首页> 外文期刊>Cell and Tissue Research >Characterization of germ cells from pre-pubertal bull calves in preparation for germ cell transplantation
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Characterization of germ cells from pre-pubertal bull calves in preparation for germ cell transplantation

机译：青春期前公牛犊生殖细胞的特征为生殖细胞移植做准备

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Although methods to assess testis cell populations are established in mice, the detailed validation of similar methods for bovine testis cells is necessary for the development of emerging technologies such as male germ cell transplantation. As young calves provide donor cells for germ cell transplantation, we characterized cell populations from three key pre-pubertal stages. Nine Angus bull calves were selected to represent three stages of testis development at ages (and testis weights) of 2–3 months (Stage 1, 10 g), 4–5 months (Stage 2, 35 g), and 6–7 months (Stage 3, 70 g). The proportion and absolute numbers of germ and somatic cells in fixed sections and from enzymatically dissociated seminiferous tubules were assessed. Germ cells were identified by DBA and PGP9.5 staining, and Sertoli cells by vimentin and GATA-4 staining. Staining of serial sections confirmed that DBA and PGP9.5 identified similar cells, which were complementary to those stained for vimentin and GATA-4. In fixed tubules, the proportion of cells within tubules that were positive for DBA and PGP9.5 increased nearly three-fold from Stage 1 to Stage 2 with no further increase at Stage 3. Absolute numbers of spermatogonia also increased between Stages 1 and 2. After enzymatic dissociation of tubules, three times more DBA- and PGP9.5-positive cells were isolated from Stage 3 testes than from either Stage 1 or 2 testes. A higher proportion of spermatogonia was observed after enzymatic isolation than were present in seminiferous tubules. These data should help to predict the yield and expected proportions of spermatogonia from three distinct stages of testis development in pre-pubertal bull calves.
机译：尽管在小鼠中建立了评估睾丸细胞数量的方法，但是对于牛睾丸细胞类似方法的详细验证对于开发新兴技术（例如雄性生殖细胞移植）是必要的。由于幼小牛为生殖细胞移植提供了供体细胞，因此我们表征了三个关键的青春期前阶段的细胞群体。选择了九只安格斯公牛犊来代表在2–3个月（第1阶段，10克），4-5个月（第2阶段，35克）和6-7个月的年龄（和睾丸重量）的三个阶段的睾丸发育（阶段3，70克）。评估了固定切片和酶解的生精小管中生殖细胞和体细胞的比例和绝对数量。通过DBA和PGP9.5染色鉴定生殖细胞，通过波形蛋白和GATA-4染色鉴定支持细胞。连续切片的染色证实，DBA和PGP9.5鉴定出相似的细胞，与波形蛋白和GATA-4染色的细胞互补。在固定小管中，从第1阶段到第2阶段，DBA和PGP9.5阳性的小管内细胞比例增加了近三倍，而在第3阶段则没有进一步增加。在第1阶段和第2阶段之间，精子的绝对数量也增加了。肾小管酶解后，从阶段3的睾丸中分离出的DBA和PGP9.5阳性细胞的数量是从阶段1或2的三倍。酶分离后观察到的精原细胞比例高于生精小管中的精原细胞比例。这些数据应有助于预测青春期前公牛犊睾丸发育三个不同阶段精原细胞的产量和预期比例。

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