免疫磁珠分选兔前软骨干细胞及细胞研究平台的建立

发布时间：2018-05-01 12:25

本文选题：前软骨干细胞 + 免疫磁珠分选　；参考：《中国矫形外科杂志》2013年13期

【摘要】：[目的]研究兔前软骨干细胞PSCs,(precartilagious stem cells)的分离、培养方法及增殖、表型特征,为细胞移植治疗椎间盘退变,提供合适的种子细胞。[方法]取胎兔骨骺周围软骨膜(LaCroix环)中的细胞进行原代培养,然后采用免疫磁珠分离系统分选纯化PSCs。体外培养,传代,冻存复苏,绘制生长曲线,观察细胞特性。分别采用免疫组化、免疫荧光、RT-PCR等方法鉴定纯化后的PSCs。[结果]免疫磁珠分离可获得纯度较高的兔PSCs,培养后成活率高,细胞状态良好。细胞冻存复苏后,细胞增殖速度、细胞形态及表面标志物无明显变化。免疫组化、免疫荧光、RT-PCR等方法鉴定后,细胞都有明显的PSCs表面特异性标记物的表达。[结论]PSCs存在于LaCroix环中,免疫磁珠分离法得到的PSCs,体外培养条件下,细胞增殖较快,生物学特性稳定。
[Abstract]:[objective] to study the isolation, culture, proliferation and phenotypic characteristics of rabbit anterior chondrocyte stem cells (PSCs) in order to provide suitable seed cells for the treatment of disc degeneration. [methods] the cells from the periepiphyseal perichondrium of fetal rabbits were cultured in primary culture, and PSCs were purified by immunomagnetic bead separation system. Culture in vitro, passage, cryopreservation and resuscitation, draw growth curve, observe cell characteristics. The purified PSCswere identified by immunohistochemistry and immunofluorescence reverse transcription polymerase chain reaction (RT-PCR). [results] PSCs with high purity could be obtained by immunomagnetic beads separation. The survival rate was high and the cells were in good condition. After cryopreservation and resuscitation, there were no significant changes in cell proliferation rate, cell morphology and surface markers. The expression of PSCs surface specific markers was detected by immunohistochemistry and RT-PCR. [conclusion] PSCs exists in the LaCroix ring, and the PSCs obtained by immunomagnetic beads method can proliferate rapidly and have stable biological characteristics in vitro.
【作者单位】：南京医科大学附属无锡市人民医院骨科;
【基金】：南京医科大学科技发展基金重点项目(项目编号:2010NJMUZ64,No.2011NJMU205) 国家自然科学基金青年科学基金项目(项目编号:81101374)
【分类号】：R329

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