天津农学院学报 ›› 2026, Vol. 33 ›› Issue (2): 57-62.doi: 10.19640/j.cnki.jtau.2026.02.010

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不同供体年龄犬骨髓间充质干细胞体外增殖与分化能力的比较研究

王欣欣1, 李珂萱1, 白春娜2, 韩厚玉2, 葛秀国1, 李吉霞1,通信作者   

  1. 1.天津农学院 动物科学与动物医学学院,天津 300392;
    2.百欧派(天津)生物技术有限公司,天津 300350
  • 收稿日期:2024-08-01 出版日期:2026-04-30 发布日期:2026-05-06
  • 通讯作者: 李吉霞(1977—),女,副教授,博士,研究方向为细胞与分子生物学。E-mail:jixiali2002@163.com。
  • 作者简介:王欣欣(1998—),女,硕士在读,研究方向为动物医学。E-mail:2821988066@qq.com。
  • 基金资助:
    天津市农业动物繁殖与健康养殖重点实验室开放课题基金(无编号)

Comparison of the proliferation and differentiation ability in vitro of canine bone marrow mesenchymal stem cells from donors of different ages

Wang Xinxin1, Li Kexuan1, Bai Chunna2, Han Houyü2, Ge Xiuguo1, Li Jixia1,Corresponding Author   

  1. 1. College of Animal Science and Veterinary Medicine, Tianjin Agricultural University, Tianjin 300392, China;
    2. Biopet (Tianjin)Biotechnology Co., Ltd., Tianjin 300350, China
  • Received:2024-08-01 Online:2026-04-30 Published:2026-05-06

摘要: 骨髓间充质干细胞(BMSCs)是一类存在于骨髓组织中、具有多向分化潜力的成体干细胞。近20年来,已有大量科学研究证明了BMSCs在疾病治疗中的潜力。然而,关于BMSCs供体来源对细胞生物学特性和细胞移植效果的影响仍有待研究。为了探索年龄对犬BMSCs增殖和分化能力的影响,本试验采用同种方法分离和培养来源于新生仔犬和青年犬的BMSCs,并对两种来源的BMSCs进行比较。结果发现,两种BMSCs在分离培养6 d都形成了较密集的细胞集落,但细胞生长形态存在差异,新生仔犬骨髓间充质干细胞(BMSCs-N)形态较细长,呈纤维型,而青年犬骨髓间充质干细胞(BMSCs-Y)呈梭型。细胞增殖试验表明,BMSCs-N组群体倍增时间((31.99±0.26)h)显著少于BMSCs-Y组群体倍增时间((34.66±1.15)h)。细胞划痕试验显示,细胞损伤9 h后,BMSCs-N组的愈合速率((45.51±1.58)%)显著高于BMSCs-Y组((37.62±2.45)%);损伤24 h,BMSCs-N组划痕愈合率为(72.13±0.97)%,BMSCs-Y组为(50.28±3.53)%。体外分化能力比较发现,BMSCs-N组成脂细胞分化率((34.25±0.22)%)高于BMSCs-Y组((25.23±1.82)%,P<0.01),BMSCs-N组成骨细胞分化率((31.34±3.88)%)高于BMSCs-Y组((15.54±0.65)%,P<0.01)。利用qRT-PCR检测多能性基因mRNA表达水平,结果显示BMSCs-N组的Sox2、Oct-4Nanog表达水平均显著高于BMSCs-Y组(P<0.05)。试验发现,与BMSCs-Y相比,BMSCs-N体外增殖活力更加旺盛,体外分化能力也较强,研究结果为利用BMSCs进行细胞治疗提供参考。

关键词: 骨髓间充质干细胞, 增殖, 分化, 年龄,

Abstract: Bone marrow Mesenchymal stem cells(BMSCs)are a group of adult stem cells that exist in bone marrow tissue and have the potential to differentiate in multiple directions. In recent 20 years, a lot of scientific researches have proved the potential of BMSCs in the treatment of diseases. However, the effects of BMSCs donor sources on cell biological properties and transplantation efficacy remain to be investigated. In order to explore the effects of ages on the proliferation and differentiation, BMSCs from newborn and young dogs were isolated and cultured by the same method, and the BMSCs from two sources were compared. The results showed that both BMSCs could form dense cell colonies after 6 days of isolation and culture, but the growth morphology of the cells was different. BMSCs of newborn puppies(BMSCs-n)was longer and more fibrous, while BMSCs of young dogs(BMSCs-Y)was spindle-shaped. The population doubling time of BMSCs-N group(31.99±0.26)h was significantly shorter than that of BMSCs-Y group(34.66±1.15) h. The results of cell scratch test showed that the healing rate of BMSCs-N group was significantly higher than that of BMSCs-Y group((45.51±1.58)% vs.(37.62±2.45)%, P<0.05)at 9 h after injury, and that of BMSCs-N group was(72.13±0.97)% at 24 h after injury, BMSCs-Y group was(50.28±3.53)%. Compared with BMSCs-Y group, the differentiation rate of adipocytes in BMSCs-N group was higher((34.25±0.22)% vs. (25.23±1.82)%, P<0.01), and that of osteoblasts in BMSCs-N group was higher than that of BMSCs-Y group((31.34±3.88)% vs. (15.54±0.65)%, P<0.01). The mRNA expression levels of pluripotency genes in BMSCs-N group were significantly higher than those in BMSCs-Y group(P<0.05). It was found that BMSCs-N were more proliferative and differentiated in vitro than BMSCs-Y. The results of this study may provide a reference for the cell therapy by BMSCs.

Key words: bone marrow mesenchymal stem cells, proliferation, differentiation, age, canine

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