Adipose tissue is an ideal stem cell source for tissue reconstruction of bone，cartilage，and soft tissue defects

目的 验证人脂肪基质细胞是否具有向成骨细胞、软骨细胞、脂肪细胞分化的能力,从而为骨、软骨、软组织再建寻找一种理想的干细胞来源.方法 分别用成骨向分化培养基（DMEM＋10?S＋地塞米松＋维生素C＋β-甘油磷酸）、软骨向分化培养基（DMEM＋1?S＋胰岛素＋维生素C＋转化生长因子β1）及脂肪向分化培养基（DMEM＋10?S＋地塞米松＋胰岛素＋吲哚美辛＋异丁基甲基黄嘌呤）诱导人脂肪基质细胞向成骨细胞、软骨细胞及脂肪细胞分化.用von Kossa和碱性磷酸酶染色鉴定成骨细胞分化,而软骨细胞分化和脂肪细胞分化分别用Alcian blue染色和油红O染色显示.成骨细胞、软骨细胞以及脂肪细胞特异相关或标志基因的表达用RT-PCR检测.结果 体外实验表明,人脂肪基质细胞在定向分化诱导剂的作用下可分别向成骨细胞、软骨细胞及脂肪细胞分化.结论 人脂肪基质细胞中包含有多向分化能力的干细胞,可用于今后骨、软骨、软组织的组织工程再建.

Adipose tissue is an ideal stem cell source for tissue reconstruction of bone, cartilage, and soft tissue defects

Background The use of stem cells for bone, cartilage, and adipose tissue engineering is promising for hard and soft tissue reconstruction. Bone marrow contains several cell populations, including mesenchymal stem cells (MSCs) that are capable of differentiating into adipogenic, osteogenic and chondrogenic cells. However, harvesting of stem cells from bone marrow has potential limitations such as low cell yield, patient discomfort and high cost. Consequently, alternative sources of autologous adult stem cells, obtainable in large quantities,under local anesthesia and with minimal patient discomfort are currently under investigation. Human adipose tissue-derived stromal cells (hADSCs), may be one such potential source as large quantities can be easily obtained by liposuction. Objective This study aims to verify whether hADSCs can differentiate along an osteoblastic, chondrocytic and adipocytic lineage, which could then potentially be used to restore bone, cartilage and soft tissue defects, respectively. Methods Osteogenic, chondrogenic, and adipogenic differentiation of hADSCs were induced by osteogenic medium (Dulbecco's modified Eagle's medium + 10% fetal bovine serum + dexamethasone + ascorbate + β-glycerophosphate), chondrogenic medium (Dulbecco's modified Eagle's medium + 1% fetal bovine serum + insulin + ascorbate + transforming growth factor-β1), or adipogenic medium(Dulbecco's modified Eagle's medium + 10% fetal bovine serum + dexamethasone + insulin + indomethacin + isobutyl-methylxanthine ) respectively for 1 ～ 3 weeks. Osteogenic differentiation was assessed by yon Kossa and alkaline phosphatase staining, while chondrogenic and adipogenic differentiation were assessed by Alcian blue staining and Oil Red O staining respectively. Expression of osteoblast specific genes, chondrocyte specific genes, and adipocyte specific genes were confirmed by RT-PCR.Results In the presence of lineage-specific induction factors, hADSCs differentiated in vitro into osteogenic, chondrogenic, and adipogenic cells. Conclusions hADSCs contain multipotent cells and may represent an ideal stem cell source for use in hard and soft tissue engineering.