Liu Feng, Cai Chunhong, Wu Xin, Cheng Yanxia, Lin Tao, Wei Guanghui, He Dawei
Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
Department of Urinary Surgery, Children's Hospital, Ministry of Education Key Laboratory of Child Development and Disorder, Key Laboratory of Pediatrics in Chongqing (CSTC2009CA5002), Chongqing Medical University, Chongqing, China.
Theriogenology. 2016 Jan 15;85(2):193-9. doi: 10.1016/j.theriogenology.2015.09.012. Epub 2015 Sep 10.
To compare KnockOut serum replacement (KSR) and fetal bovine serum (FBS) for the development of germ cells. Testicular tissues from Sprague-Dawley rats were cultured for 4 weeks in culture media supplemented with FBS or KSR. Tissue area was measured at the beginning and end of the culturing period. Testicular histology, development of the germ cells, and the diameter of seminiferous tubules were analyzed by hematoxylin and eosin staining. After 4 weeks in culture, apoptosis and expression of the stage-specific spermatogenesis marker genes Kit, Sycp3, and Crisp1 were assayed. Tissues cultured in KSR-supplemented media were able to sustain growth and gradually increase seminiferous tubule diameter during the culture period. In addition, spermatogonia, primary spermatocytes, secondary spermatocytes, and round spermatids were observed after 4 weeks in culture, and reverse transcription-PCR confirmed expression of the marker genes. In comparison, tissues cultured in FBS-supplemented media showed dwindling testicular organization, necrotic seminiferous tubules, and expression of Kit, but inconsistent expression of Sycp3 and Crisp1 KnockOut serum replacement outperforms FBS as a growth media supplements for culturing immature spermatogonial tissue culture.
比较KnockOut血清替代物（KSR）和胎牛血清（FBS）在生殖细胞发育中的作用。将来自Sprague-Dawley大鼠的睾丸组织在补充有FBS或KSR的培养基中培养4周。在培养期开始和结束时测量组织面积。通过苏木精和伊红染色分析睾丸组织学、生殖细胞发育以及生精小管直径。培养4周后，检测凋亡情况以及阶段特异性生精标记基因Kit、Sycp3和Crisp1的表达。在补充有KSR的培养基中培养的组织在培养期间能够维持生长并逐渐增加生精小管直径。此外，培养4周后观察到精原细胞、初级精母细胞、次级精母细胞和圆形精子细胞，逆转录-聚合酶链反应证实了标记基因的表达。相比之下，在补充有FBS的培养基中培养的组织显示睾丸组织结构逐渐减少、生精小管坏死以及Kit的表达，但Sycp3和Crisp1的表达不一致。作为培养未成熟精原细胞组织培养的生长培养基补充物，KnockOut血清替代物优于胎牛血清。
