CAJ | 학술논문

目的探讨新型无血清培养基 SYL-SF 对脐带间充质干细胞的体外培养和扩增作用. 方法机械分离方法获得脐带间充质干细胞,从 P1代开始将含小牛血清的原代培养基 SYL-NBCS 更换为实验组SYL-SF 和对照组 MSCM-SF 两种无血清培养基对细胞进行培养、传代、扩增和维持.利用 Alexa Fluor 488 annexin V检测细胞凋亡率, CCK-8法比较传代细胞的增殖效率,并对细胞进行形态、表型、染色体核型和分化能力等方面的检测和评价. 结果分离得到的单个脐带间充质干细胞在原代培养基SYL-NBCS 中培养8~15 d 可获得原代(P0)细胞,更换为无血清培养基后,SYL-SF 组细胞凋亡率明显低于MSCM-SF 组;在细胞传代过程中 SYL-SF 组细胞增殖能力较强且维持 MSCs 特异性表面标志和向脂肪、骨、软骨等多向分化的潜能,并且在传代过程中维持正常染色体核型. 结论 SYL-SF 无血清培养基可以对 UC-MSCs 进行良好地体外扩增和维持.
목적탐토신형무혈청배양기 SYL-SF 대제대간충질간세포적체외배양화확증작용. 방법궤계분리방법획득제대간충질간세포,종 P1대개시장함소우혈청적원대배양기 SYL-NBCS 경환위실험조SYL-SF 화대조조 MSCM-SF 량충무혈청배양기대세포진행배양、전대、확증화유지.이용 Alexa Fluor 488 annexin V검측세포조망솔, CCK-8법비교전대세포적증식효솔,병대세포진행형태、표형、염색체핵형화분화능력등방면적검측화평개. 결과분리득도적단개제대간충질간세포재원대배양기SYL-NBCS 중배양8~15 d 가획득원대(P0)세포,경환위무혈청배양기후,SYL-SF 조세포조망솔명현저우MSCM-SF 조;재세포전대과정중 SYL-SF 조세포증식능력교강차유지 MSCs 특이성표면표지화향지방、골、연골등다향분화적잠능,병차재전대과정중유지정상염색체핵형. 결론 SYL-SF 무혈청배양기가이대 UC-MSCs 진행량호지체외확증화유지.
Objective To investigate the effect of a new serum-free medium SYL-SF on the culture and expansion of umbilical cord derived mesenchymal stem cells (UC-MSCs) . @@@@Methods UC-MSCs were obtained from aborted human umbilical cord by mechanical isolation method, and were maintained in primary culture medium SYL-NBCS. From passage 1 (P1), UC-MSCs were maintained and expanded in two different types of serum-free medium SYL-SF and MSCM-SF. Apoptosis was analyzed with Alexa Fluor 488 annexin V apotosis kit and cellular proliferation was analyzed with CCK-8 method. Cellular morphology, karyotype, cell surface marker and differentiation capacity were also detected. @@@@Results Primary UC-MSCs (P0) are obtained 8 to 15 days after the mechanical isolation and cultured in primary medium SYL-NBCS. When the cells are changed into serum-free medium, the apoptosis rate of the cells cultured in SYL-SF is significantly lower than those in MSCM-SF. In SYL-SF medium, UC-MSCs expend faster and maintain MSCs-specific surface markers as well as the multipotent differentiation potential towards fat, bone and cartilage. They also maintain normal karyotype after 8 passages of culture in SYL-SF. @@@@Conclusion SYL-SF as a novel serum-free medium can be used to support the expansion and maintenance of UC-MSCs in vitro.