畜牧与兽医2026，Vol.58Issue(9)：86-95,10.
渠溪猪肺源间充质干细胞体外培养与生物学特性研究
Study on the in vitro culture and biological characteristics of mesenchymal stem cells from the lungs of Quxi pigs
摘要
Abstract
The aim of this study was to isolate and culture pulmonary mesenchymal stem cells(PMSCs)from the lungs of Quxi pigs in vitro,and to investigate their biological characteristics,proliferation and differentiation capabilities.Lung tissues were aseptically isolated from 44-day-old healthy Chuxi pigs,and PMSCs were cultured in vitro using the tissue block adhesion method.In this study,cells from the 3rd,15th,and 25th generations were selected for detection and analysis.The population doubling time was calculated by drawing the cell growth curve,and the colony formation ability was evaluated.The scratch assay was used to simulate the wound healing process in vivo to as-sess the migration activity of the cells.Chromosome karyotype analysis was employed to verify the genetic stability of the cells.The multi-directional differentiation potential was identified through osteogenic,adipogenic,and chondrogenic differentiation experiments,combined with specific staining and semi-quantitative PCR techniques.Meanwhile,the expression levels of typical surface markers of mesenchymal stem cells were detected by immunofluorescence staining and semi-quantitative PCR.The results showed that the isolated PMSCs were long spindle-shaped and grew in parallel,with a growth curve in an"S"shape.The population doubling time of the 15th generation was between that of the 3rd and 25th generations,and the colony formation rate decreased with increasing passage number.The chromosome karyotype analysis revealed a normal diploid karyotype(2n=38).Immunofluorescence and semi-quantitative PCR techniques detected positive reac-tions for CD29,CD73,CD90,and CD105 markers on the surface of PMSCs,while CD34 was negative,which met the identification criteria of mesenchymal stem cells.Additionally,the results of the induction differentiation experiments confirmed that PMSCs could successfully dif-ferentiate into osteoblasts,adipocytes,and chondrocytes.In conclusion,this study successfully isolated and cultured PMSCs with typical mesenchymal stem cell characteristics from the lung tissues of Chuxi pigs,demonstrating their multi-directional differentiation potential and good genetic stability.This cell line provides a basis for preservation of genetic resources and also offers potential available resources for cell transplantation.关键词
渠溪猪/肺间充质干细胞/体外分离/培养鉴定/诱导分化Key words
Quxi pig/pulmonary mesenchymal stem cells/in vitro isolation/culture and identification/induced differentiation分类
农业科技引用本文复制引用
张婷婷,闫小妮,王丰璇,白娜,李楠,关伟军..渠溪猪肺源间充质干细胞体外培养与生物学特性研究[J].畜牧与兽医,2026,58(9):86-95,10.基金项目
国家重点研发计划项目(2021YFD1200303) （2021YFD1200303）
国家家养动物种质资源平台项目(2026年) （2026年）
中国农业科学院科技创新工程项目(ASTIP-IAS01) （ASTIP-IAS01）
黑龙江省自然科学基金项目(PL2024H013) （PL2024H013）
