中国实用口腔科杂志

中国实用口腔科杂志

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人脂肪组织来源血管周干细胞成骨、成脂、成软骨分化能力研究

张蕾，孟芝竹，张斌   

  1. 中国医科大学口腔医学院·附属口腔医院口腔颌面外科，辽宁 沈阳  110002
  • 出版日期:2018-07-15 发布日期:2018-11-08
  • 作者简介:中国医科大学口腔医学院青年科研启动基金（K101593-18-05）

  • Online:2018-07-15 Published:2018-11-08

摘要：

目的    研究从人脂肪组织中提取的血管周干细胞（human perivascular stem cells，hPSCs）的特点，并探讨其成骨、成脂及成软骨分化的能力。方法    采用流式荧光细胞分选技术（FACS）在12例行吸脂手术患者的脂肪标本中分选出人基质血管成分（human stromal vascular fraction，hSVF）和由周皮细胞（CD34-、CD146+、CD45-）与外膜细胞（CD34+、CD146-、CD45-）组成的hPSCs进行培养，比较其克隆增殖能力，然后将2种细胞进行成骨、成脂和成软骨诱导，诱导结束后分别进行茜素红染色、油红O染色和阿尔新蓝染色，并检测成骨诱导后的成骨相关基因mRNA表达。结果    hSVF和hPSCs均以纺锤形成纤维样细胞生长，hPSCs呈现出更快的融合趋势，且细胞形态更均一。hPSCs细胞相比于hSVF具有更强的克隆增殖能力（P < 0.05）。hPSCs细胞在成骨及成软骨方向比hSVF具有更强优势，而hSVF在成脂方向比hPSCs具有更强优势。成骨诱导后，hPSCs的成骨基因碱性磷酸酶（alkaline phosphatase，ALP）和骨钙素（osteocalcin，OCN）的相对表达量要明显高于hSVF（P < 0.05）。结论    hPSCs细胞具有干细胞的多项分化潜能，且其在成骨方向具有很强优势，可成为骨组织工程学中理想的种子细胞来源。

关键词: 人血管周干细胞, 人基质血管成分, 成骨分化, 成脂分化, 成软骨分化

Abstract:

Objective    To extract human perivascular stem cells（hPSCs）from fat tissue，and then to detect its abilities of osteogenic differentiation，adipogenic differentiation，and chondrogenic differentiation. Methods     Human stromal vascular fraction（hSVF）and hPSCs were purified from 12 samples of human lipoaspirate by FACS. After cell culture， the proliferation abilities of hSVF and hPSCs were compared. Then two kinds of cells were induced into osteogenic differentiation，adipogenic differentiation，and chondrogenic differentiation. Results were obtained by alizarin red staining，oil red staining，alcian blue staining，and qRT-PCR. The mRNA expression of osteogenesis-related genes were detected. Results    hSVF and hPSCs grew like spindle fibroblast. hPSCs fused faster than hSVFs，and had homogeneous morphology. hPSCs had stronger cloning and proliferation ability than hSVF（P < 0.05）. hPSCs had stronger osteogenic differentiation ability and chondrogenic differentiation ability than hSVF（P < 0.05），while hSVF had stronger adipogenic differentiation ability than hPSCs. Alkaline phosphatase（ALP）and osteocalcin（OCN）had higher expression in hPSCs than in hSVF after osteogenesis（P < 0.05）. Conclusion    hPSCs have the multipotential differentiation ability as stem cells，and have strong advantages in the direction of osteogenesis. hPSCs can be an ideal source of seed cells in bone tissue engineering.

Key words: human perivascular stem cells, hPSCs；human stromal vascular fraction, hSVF；osteogenic differentiation；adipogenic differentiation；chondrogenic differentiation