China Animal Husbandry & Veterinary Medicine ›› 2025, Vol. 52 ›› Issue (6): 2519-2530.doi: 10.16431/j.cnki.1671-7236.2025.06.006

• Physiological and Biochemical • Previous Articles    

Study on Isolation Culture and Biological Characteristics of Pancreatic Mesenchymal Stem Cells in Simmental Cattle

WANG Zhongfa1,2, LIU Yanchen2, YAN Yan2, LI Minjuan1,2, HE Yunan2, GUAN Weijun2, LIU Wenzhong1   

  1. 1. College of Animal Science, Shanxi Agricultural University, Taigu 030801, China;
    2. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2025-01-03 Published:2025-05-27

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Abstract: 【Objective】 The purpose of this study was to isolate and culture pancreatic mesenchymal stem cells (PMSCs) in Simmental cattle,establish PMSCs line in Simmental cattle,study its biological characteristics,and provide reference for the preservation of germplasm resources in Simmental cattle.【Method】 PMSCs were isolated and cultured from pancreas tissue in Simmental cattle by enzyme digestion and tissue block adhesion method,and the biological characteristics of PMSCs were identified.The expressions of cell surface markers (CD34,CD73,CD90 and CD166) were measured by immunofluorescence and semi-quantitative PCR.The cell proliferation ability was determined by drawing cell growth curves of P4,P10 and P16 generations and calculating population doubling time.The cell cloning rate was determined by cell cloning test,and the cell migration rate was calculated through scratch test.The purified cells were induced into osteoblasts,adipocytes and chondrocytes respectively to detect the multipotent differentiation ability of PMSCs,and the cell genetic stability was detected by chromosome karyotype analysis test.【Result】 PMSCs could be cultured using tissue block adhesion method and passaged to the third generation for basic purification.Semi-quantitative PCR results showed that the surface markers (CD73,CD90 and CD166) were positive,but CD34,a surface antigen related to hematopoietic cells,was negative.The growth curve of PMSCs in Simmental cattle showed a typical S curve,the doubling time of P4,P10 and P16 generation cells were 40.22,43.09 and 44.50 h,and the colony formation rate were 26.25%,19.00% and 13.50%,respectively.The migration rate of the third passage cells was 44.44% by scratch test.The clear lipid droplets,cartilage masses and calcium nodules were observed after differentiation of PMSCs in vitro.Karyotype analysis results showed that PMSCs in Simmental cattle were normal diploid karyotype (2n=60,XY),and the chromosome set did not change.【Conclusion】 PMSCs isolated and cultured from pancreas tissue in Simmental cattle could stably inherit and strongly proliferate,and had the potential to differentiate into osteoblasts,adipocytes and chondrocytes.The results could provide new method for tissue repair and seed cells for the protection and preservation of germplasm resources in Simmental cattle.

Key words: Simmental cattle; pancreatic mesenchymal stem cells; isolation and culture; induced differentiation

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