治疗运动性损伤韧带来源间充质干细胞体外培养与生物学特性研究OA
In vitro Culture and Its Biological Characteristics of Ligament-derived Mesenchymal Stem Cells for Treating Sports-related Injuries
为系统评估韧带来源间充质干细胞(ligament-derived mesenchymal stem cells,LMSCs)的关键生物学潜能,以验证其加速运动性损伤修复的可行性,为相关治疗提供新思路,选取23 d龄北京鸭胎鸭骨骼肌韧带为研究对象,利用Ⅳ型胶原酶消化法和贴壁培养法在体外进行LMSCs的多代次培养.按照国际公认的细胞质量检测项目和标准对LMSCs的形态、活率、表面标志物、免疫组织化学、多向诱导分化潜能、阳性率、核型、微生物污染、迁移能力、克隆能力、生长曲线、群体倍增时间、冻存复苏后形态与活率等20项指标进行检测.结果表明,体外培养的LMSCs可传至P31代,其细胞形态呈长梭形,旋涡状生长.随着代次增高,细胞生长速度逐渐缓慢,活力渐弱,传代间隔延长.流式细胞术结果显示,培养的LMSCs具有很高的细胞纯度.RT-PCR和免疫荧光结果显示,Collagen Ⅰ、Collagen Ⅲ、CD73、CD44和Tenascin-C均呈阳性表达.诱导分化结果显示,LMSCs具有在体外诱导分化为多种类型组织来源体细胞的潜能.染色体核型分析显示,LMSCs染色体数目正常,无染色体畸变.综上,利用北京鸭这一我国重要的畜禽遗传资源建立了用于治疗运动性损伤LMSCs体外培养的科学体系,为畜禽遗传资源的细胞水平保存与功能评价提供了技术支撑.
To systematically evaluate the key biological potential of ligament-derived mesenchymal stem cells(LMSCs)and validate their feasibility in accelerating the repair of sports injuries,providing new insights for related therapies,the ligaments from skeletal muscles of 23-day-old Beijing duck embryos were as the research subject,and the type IV collagenase digestion and adhesion-based culture methods were used to perform multi-pass in vitro culture of LMSCs.Following internationally recognized cell quality assessment protocols and standards,the LMSCs underwent morphological observation,viability testing,surface marker analysis,immunohistochemistry,multi-lineage differentiation potential(adipogenic,osteogenic and chondrogenic differentiation capabilities),positive rate,karyotype,microbial contamination(bacteria,fungi,protozoa,mycoplasma and pathogens),migration capacity,cloning ability,growth curve plotting,doubling time,morphology and viability after cryopreservation and thawing,among 20 indexes.The results indicated that the cultured LMSCs in vitro could be passaged up to P31,exhibiting a long spindle-shaped morphology with swirling growth patterns.With increasing passage numbers,cell growth rate gradually slowed,viability diminished,and passage intervals lengthened.Flow cytometry revealed high cellular purity in cultured LMSCs.RT-PCR and immunofluorescence results confirmed positive expression of CollagenⅠ,Collagen Ⅲ,CD73,CD44 and Tenascin-C.Induction differentiation results demonstrated that LMSCs possessed the potential to differentiate in vitro into various types of tissue-derived somatic cells.Chromosome karyotyping analysis revealed normal chromosome number with no chromosomal aberrations.To sum up,a scientific system was established for in vitro culture of LMSCs for treating sports injuries,using the Beijing duck-an important livestock genetic resource in China,which provided technical support for the cellular-level preservation and functional evaluation of livestock genetic resources.
杨雪松;王丰璇;王仲发;李旻娟;张婷婷;何玉楠;袁子奥;关伟军
首都体育学院武术与表演学院,北京 100191首都体育学院武术与表演学院,北京 100191中国农业科学院北京畜牧兽医研究所,北京 100193中国农业科学院北京畜牧兽医研究所,北京 100193中国农业科学院北京畜牧兽医研究所,北京 100193中国农业科学院北京畜牧兽医研究所,北京 100193哈尔滨师范大学生命科学与技术学院,哈尔滨 150028中国农业科学院北京畜牧兽医研究所,北京 100193
生物科学
间充质干细胞体外培养与鉴定运动性损伤
mesenchymal stem cellsin vitro culture and identificationsports injury
《中国农业科技导报》 2026 (8)
164-174,11
国家重点研发计划项目(2021YFD1200303)国家自然科技资源平台项目(NGCDAR-IAS202401).
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