CAJ | 학술논문

目的从昆明鼠睾丸中克隆Bmi1基因,构建真核表达载体,并转染支持细胞,以便用作培养精原干细胞（SSCs）的滋养层.方法以5日龄昆明鼠为材料,提取小鼠睾丸组织中总RNA后,以RT-PCR技术克隆小鼠睾丸Bmi1基因,构建真核表达载体,并转染TM4细胞（睾丸支持细胞株）,在转染后40 h进行免疫荧光鉴定.结果成功克隆小鼠睾丸Bmi1基因的cDNA,测序正确;免疫荧光细胞染色显示,转染后的支持细胞中有Bmi1蛋白表达.结论本研究为以转染了Bmi1基因的支持细胞作饲养层培养SSCs奠定了基础.
목적종곤명서고환중극륭Bmi1기인,구건진핵표체재체,병전염지지세포,이편용작배양정원간세포（SSCs）적자양층.방법이5일령곤명서위재료,제취소서고환조직중총RNA후,이RT-PCR기술극륭소서고환Bmi1기인,구건진핵표체재체,병전염TM4세포（고환지지세포주）,재전염후40 h진행면역형광감정.결과성공극륭소서고환Bmi1기인적cDNA,측서정학;면역형광세포염색현시,전염후적지지세포중유Bmi1단백표체.결론본연구위이전염료Bmi1기인적지지세포작사양층배양SSCs전정료기출.
Objecave：To clone Bmi1 gene from Kunming mouse testis,construct the eukaryotic expression vector and transfect this vector into Sertoli cells in order to use the Bmi1-transfected Sertoli cells as the feeder layer to cultivate sperrnatogonial stem cells（SSCs）.Methods：Total RNA was extracted from the testes of 5 day old Kunming mice and Bmi1 was cloned and amplified using RT-PCR,inserted into the eukaryotic expression vector and transfected into sertoli cells（TM4 cell line）.Immunofluorescence with anti-Bmi1 antibodies was performed at 40 h following the transfection.Results：Bmi1 DNA was cloned successfully,and Bmi1 expressed at 40 h after transfected into Sertoli cells.Conclusion：The present study provides a basis for culturing SSCs with Bmi1-transfected sertoli cells as the feeder layer.