北京大学学报（医学版）2012，Vol.44Issue(3)：475-480,6.DOI:10.3969/j.issn.1671-167X.2012.03.028
重组人肿瘤坏死因子α对人脂肪基质细胞体外成骨向分化的影响
Recombinant human tumor necrosis factor-alpha promotes human adipose-derived stromal cells transforming into osteoblast in vitro
摘要
Abstract
Objective: To investigate the effect of recombinant human tumor necrosis factor alpha (rhTNF-a)on the osteogenesis potential of the osteo-induced human adipose-derived stromal cells (hASCs) in vitro. Methods: hASCs at passage 4 were divided into four groups according to culturing conditions; basal medium [BM, DMEM + 10% FBS + antibiotics], BM with 10 μg/L rhTNF-α, os-teogenic medium (OM, BM + dexamethasone + L-ascorbate + p-glycerophosphate) and OM with 10 μg/L rhTNF-α. On days 3,7, 14 and 21, alkaline phosphatase (ALP) activities were examined. On days 14 and 21, the staining and quantitation of calcium deposition were performed. For the cells under osteogenic induction, osteoblast- related genes, such as core-binding factor αl (Cbfal) , Osterix (Osx) and osteocalcin (OC) were analyzed with reverse transcription PCR on days 3, 7, 14, and 21, and real time PCR was performed to confirm the effect of rhTNF-α on genes expression on day 3 . Results: rhTNF-apromoted ALP activities of induced hASCs on day 14 (3. 527 ± 0. 415 vs. 2. 345 ± 0. 354, P<0.01) and on day 21 (3.106 ± 0. 105 vs. 2.442 ± 0. 163,P <0.01) and promoted calcium deposition of induced hASCs on day 14 (2. 896 ± 0. 173 vs. 0. 679 ± 0. 173, P <0. 01) and on day 21 (2.231 ±0.233 vs. 1.729 ± 0. 229, P <0. 01). RT-PCR and Real-time PCR assays showed that rhTNF-α augmented the expression of Cbfal, Osx and OC of these cells. Conclusion; The findings indicate that 10 μg/L rhTNF-a can promote the osteogenic potential of osteogenetically induced hASCs in vitro.关键词
脂肪组织/间质细胞/骨生成/肿瘤坏死因子αKey words
Adipose tissue/Stromal cells/Osteogenesis/Tumor necrosis factor-alpha分类
医药卫生引用本文复制引用
余日月,曾百进,刘云松,周永胜..重组人肿瘤坏死因子α对人脂肪基质细胞体外成骨向分化的影响[J].北京大学学报（医学版）,2012,44(3):475-480,6.基金项目
国家自然科学基金(30901693)和北京世纪坛医院科研基金资助 （30901693）
