低强度脉冲超声对人脂肪干细胞成骨诱导影响的实验研究  

Low- intensity pulsed ultrasound enhances osteogenic medium- induced osteogenic differentiation of human adipose tissue- derived stem cells in vitro

作　　者：丁涛[1] 程力[1] 朱浩明[1] 褚亚伟[1] 金磊[1] 

机构地区：[1]江苏省无锡市人民医院骨科,无锡市清扬路299号214023

出　　处：《中国矫形外科杂志》2014年第11期1013-1017,共5页Orthopedic Journal of China

基　　金：无锡市医管中心重大协作项目(编号:YGZX1111)

摘　　要：[目的]探讨低强度脉冲超声在人脂肪干细胞成骨诱导中的影响。[方法]取手术患者腹部及腰背部处脂肪,采用胶原酶消化离心贴壁法分离培养脂肪间充质干细胞(adipose derived stem cells,ADSCs),流式细胞仪检测细胞表面标记。第6代细胞分为四组:(1)阴性对照组:ADSCs于完全培养基中培养,不接受LIPUS剌激;(2)LIPUS组:ADSCs于完全培养基中培养,接受LIPUS刺激;(3)成骨诱导组:ADSCs于成骨诱导培养基中培养;(4)LIPUS联合成骨诱导组:ADSCs在成骨诱导培养基中培养,接受LIPUS刺激。[结果]ADSCs形态传至10代时仍无变化。流式细胞仪分析第6代hADSCs细胞表面特异性标记结果显示,CD105:99.61%,CD29:97.54%,CD45:0.26%,CD44:97.4%,CD14:2.51%。LIPUS+诱导组的碱性磷酸酶(ALP)表达明显高于其他组,而单用LIPUS,并不能促进成人脂肪干细胞成骨分化。[结论]LIPUS可以促进成骨诱导培养基培养下的成人脂肪干细胞的成骨分化。[ Objective] To investigate the effect of low- intensity pulsed ultrasound （LIPUS） on osteogenic medium -in- duced osteogenic differentiation of human adipose tissue - derived stem cells （hADSCs） . [ Methods ] Human adipose tissue was obtained by liposuction or abdominal operation with patient consent and approval from the Ethics Committee. hADSCs were i- solated from the tissue by collagenase digestion and cultured using the centrifugal -adherent method. Cell phenotype was identi- fied by flow cytometry. The 6th generation cells were divided into 4 groups and cultured as follows ： （ 1 ） control group ： hADSCs cultured in growth medium without LIPUS; （2） LIPUS group： hADSCs cultured in growth medium with LIPUS; （3） osteogenic medium group： hADSCs cultured in osteogenic medium without LIPUS; （4） osteogenic medium ＋ LIPUS group： hADSCs cul- tured in osteogenic medium with LIPUS. [ Results] ADSCs maintained their cell morphology even after 10 passages. The immu- nophenotype of the 6th generation hADSCs as per flow cytometry was CD105： 99. 61%, CD29： 97.54%, CD45： 0. 26%, CD44 ： 97. 4%, CD14 ： 2. 51%. The alkaline phosphatase activity of hADSCs in the osteogenic medium ＋ LIPUS group was sig- nificantly higher than that in the other groups. However, LIPUS did not affect hADSC proliferation in the presence or absence of osteogenie medium. [ Conclusion] LIPUS has a significant effect on the osteogenic differentiation of hADSCs in the presence of osteogenic medium.

关 键 词：人脂肪干细胞 细胞培养 成骨诱导 低强度脉冲超声 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]