条件重编辑共培养技术体外培养人甲状旁腺细胞的研究  

Conditionally reprogrammed co- culturing system in human parathyroid cell culture

作　　者：阮莹[1] 侯佳宁 刘仁斌 吴珏堃[1] Ruan Ying;Hou Jianing;Liu Renbin;Wu Juekun(Department of Thyroid and Breast Surgery,the Third Affiliated Hospital,Sun Yat-sen University,Guang-zhou 510630,China;Department of Gastrointestinal Surgery,Shenzhen Hospi-tal of Peking University,Shenzhen 518036,China)

机构地区：[1]中山大学第三附属医院甲状腺乳腺外科,广州510630 [2]北京大学深圳医院胃肠外科,深圳518036

出　　处：《中华实验外科杂志》2018年第9期1641-1643,共3页Chinese Journal of Experimental Surgery

摘　　要：目的利用条件重编辑共培养（CRC）体系对人原代甲状旁腺细胞进行体外培养，建立具有生理功能且能够稳定生长的甲状旁腺主细胞株。方法使用放射后的成纤维细胞3T3-J2与Rho激酶抑制剂Y-27632（5×10μmol／L）构建CRC培养体系，人甲状旁腺原代细胞与之共培养，通过形态学、化学发光法检测甲状旁腺激素等指标评估人甲状旁腺细胞生长状况和生理活性。结果CRC培养体系中，原代甲状旁腺细胞体外培养时间为50～55d，有效时间窗为30-35d，持续传代2次，证实其具有激素内分泌活性。结论成功建立具有生理活性的甲状旁腺主细胞株，为甲状旁腺组织的进一步研究提供细胞基础。Objective To in vitro culture human parathyroid primary cells using the conditionally reprogrammed co - culture （CRC） system for establishing a parathyroid chief cell line which can grow and transfer stably in vitro. Methods The irradiated 3T3 - J2 cell line was combined with the Rho kinase in- hibitor （ Y - 27632, 5×10^-6 mol/L） and type I collagen to construct the CRC system. Human parathyroid primary cells were co - cultured in the CRC system. The growth activity and pathological features of human parathyroid cells were evaluated by methodology and parathyroid hormone concentration detected by chemi- luminescent immunoassay. Results In CRC system, the co - culture time of parathyroid chief cell line was 50 -55 days, with a significant culturing time window for 30 -35 days. Total culture generation was 3. Parathyroid hormone was detected in the medium and fluctuated according to calcium ion concentration. Conclusion A relatively stable, physiologically active parathyroid chief cell line could be built successful- ly in the CRC system.

关 键 词：甲状旁腺 体外培养 条件重编辑共培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]