雪貂脂肪间充质干细胞的分离、分化与鉴定
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(中国医学科学院医学实验动物研究所, 北京协和医学院卫生部人类疾病比较医学重点实验室干细胞与临床转化实验室,北京 100021)

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Q95-33


Isolation, characterization and differentiation of ferret adipose⁃derived stem cells
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(Laboratory of Stem Cells and Translational Medicine, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences & Comparative medical Center, Peking Union Medical College, Beijing 100021, China)

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    摘要:

    目的 分离?多向诱导分化并进行鉴定雪貂脂肪间充质干细胞?方法 无菌采取雪貂腹部皮下脂肪,机械剪碎为0. 5 mm2 , 采用含0. 075%的胶原酶I 消化,应用高糖?DMEM 培养基(添加4 ng/ mL β?FGF)培养,观察细胞形态特征,流式细胞术分析细胞表面抗原标志的表达,并检测其体外成脂?成骨?成软骨分化能力,还进一步尝试了诱导神经分化?结果 运用组织块贴壁法可以从新鲜脂肪组织中分离到贴壁生长的类似成纤维细胞样细胞,流式分析结果显示高表达CD29(53. 1%),CD90(99. 6%),CD105(93. 7%),低表达CD11b(36. 3%),CD45 (28. 3%)?这些细胞在体外经诱导可以分化成为脂肪?骨?软骨和神经元?结论 从雪貂脂肪组织中可以分离得到脂肪间充质干细胞,这些细胞具有多向分化能力?

    Abstract:

    Objective The objective of this study was to isolate, characterize, and differentiate ferret adipose?derived stem cells (ADSCs). Methods Subcutaneous fat tissues from ferret abdomens were obtained via germ?free procedures, mechanically minced into 0. 5 mm3 pieces, and digested with 0. 075% collagenase. Subsequently, ADSCs were cultured in high?glucose DMEM medium containing 10% FBS, antibiotics, and 4 ng/ mL β?FGF. Then, analyses of cell morphology and cell surface antigen expression were made by flow cytometry. Finally, osteogenic, adipogenic, chondrogenic, and neural differentiation were examined using different method. Results We isolated and identified ADSCs from fresh fat tissues of ferrets, which demonstrated a more fibroblast?like morphology. FACS analysis show high expression of CD29 (53. 1%), CD90 (99. 6%) and CD105 (93. 7%), but low expression of CD11b (36. 3%) and CD44 (28. 3%). The ADSCs could be differentiated into adipocytes, osteoblasts, chondrocytes, and neurons by in vitro induction. Conclusions ADSCs can be isolated from the subcutaneous fat tissue of ferrets, and these cells have multi?directional differentiation ability.

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石桂英,李珂雅,徐艳峰,韩云林,高舒平.雪貂脂肪间充质干细胞的分离、分化与鉴定[J].中国实验动物学报,2018,26(5):603~609.

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  • 收稿日期:2018-04-20
  • 在线发布日期: 2018-11-09
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