组织工程与重建外科杂志

组织工程与重建外科杂志 ›› 2013, Vol. 9 ›› Issue (4): 190-194.doi: 10.3969/j.issn.1673-0364.2013.04.003

• 论著 • 上一篇    下一篇

不同代次小鼠脂肪干细胞增殖能力及体外成脂能力的实验研究

蒋婷，杨泽龙，白倩，周广东，刘伟   

  1. 南充市中心医院烧伤整形美容外科；川北医学院第二临床学院；南充市中心医院骨科；上海交通大学医学院附属第九人民医院整复外科,上海市组织工程研究重点实验室
  • 发布日期:2020-07-23

Proliferation Assay and Adipogenic Ability of Mouse Adipose-derived Stem Cells on Different Passages in Vitro

JIANG Ting,YANG Zelong,BAI Qian,ZHOU Guangdong,LIU Wei   

  1. Department of Burn and Plastic Surgery,Nanchong Central Hospital,The Second Clinical College of North Sichuan Medical College; Department of Orthopedis Surgery,Nanchong Central Hospital; Department of Plastic and Reconstructive Surgery,Shanghai Ninth People’s Hospital,Shanghai Jiaotong University School of Medicine,Shanghai Key Laboratory of Tissue Engineering
  • Published:2020-07-23

摘要： 目的观察不同代次小鼠脂肪干细胞(mASCs)增殖能力及体外成脂能力,为组织工程种子细胞的合理选用提供实验依据。方法采用胶原酶消化小鼠腹股沟皮下脂肪组织,贴壁培养,体外传代扩增。实验采用原代(P0)、第2代(P2)和第5代(P5)细胞。观察不同代次mASCs的细胞形态及增殖能力;经高密度接种培养4周及成脂诱导2周,通过油红染色、RT-PCR检测,探讨其诱导成脂分化潜能及自发成脂能力。结果小鼠脂肪干细胞经传代后,外观主要呈长梭形及不规则形,不同代次的mASCs均具有很强的体外增殖能力;P0细胞具有极强的体外自发成脂能力,P2细胞自发成脂能力明显降低,至P5时基本没有体外自发成脂能力;P0、P2、P5细胞具有相似的诱导成脂分化潜能。结论随着培养代次的增加,mASCs自发成脂能力降低而诱导成脂分化潜能基本不变,扩增至第5代时仍维持了较强的增殖能力及良好的干细胞特性。在一定范围内mASCs体外传代扩增能明显减少其脂肪前体细胞的含量,有助于间充质干细胞纯化。

关键词: 小鼠, 脂肪干细胞, 不同代次, 成脂分化

Abstract: Objective To observe the proliferation capacity and adipogenic ability of mouse adipose-derived stem cells (mASCs) on different passages in vitro and to provide experimental evidence for rational selection of seed cells. Methods The mouse inguinal fat tissues were digested with collagenase, and cells were adhered to dishes, passage-cultured. The mASCs were divided into three groups: primary cells (P0), the second generation cells (P2) and the fifth generation cells (P5), and cell morphology and proliferation capacity of different passages of mASCs were observed. After four weeks of non-induction culture with high density seeding, the spontaneous adipogenic ability was observed through oil red staining and PT-PCR analysis. After two weeks of adipogenic induction, the adipogenic potential of mASCs was observed through oil red staining and PT-PCR analysis. Results The mouse adipose-derived stem cells displayed fibroblast-like morphology or irregular shape and different passages of them all had strong proliferation capacity. Primary cells had extremely strong spontaneous adipogenic ability in vitro. The spontaneous adipogenic ability of the second generation cells obviously reduced, and there was hardly any spontaneous adipogenic ability at passage 5. P0, P2 and P5 had similar induced adipogenic potential. Conclusion With the generation increasing, spontaneous adipogenic ability of mASCs reduced, but induced adipogenic potential almost unchanged. The fifth generation cells still maintain strong proliferation capacity and properties of stem cells. In a certain range of passage and amplification could reduce the content of pre-adipocyte and help purify the mesenchymal stem cells.

Key words: Mouse, Adipose-derived stem cells, Different passages, Adipogenic differentiation

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