中国农业科技导报2026，Vol.28Issue(8)：164-174,11.DOI:10.13304/j.nykjdb.2025.0746
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
In vitro Culture and Its Biological Characteristics of Ligament-derived Mesenchymal Stem Cells for Treating Sports-related Injuries
摘要
Abstract
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.关键词
间充质干细胞/体外培养与鉴定/运动性损伤Key words
mesenchymal stem cells/in vitro culture and identification/sports injury分类
生物科学引用本文复制引用
杨雪松,王丰璇,王仲发,李旻娟,张婷婷,何玉楠,袁子奥,关伟军..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[J].中国农业科技导报,2026,28(8):164-174,11.基金项目
国家重点研发计划项目(2021YFD1200303) （2021YFD1200303）
国家自然科技资源平台项目(NGCDAR-IAS202401). （NGCDAR-IAS202401）
