人脂肪间充质干细胞体外高效扩增新方法

目的 运用低强度脉冲场超声(LIPUS)刺激促进人脂肪间充质干细胞(hASCs)增殖,为临床干细胞体外高效增殖提供新方法.方法 通过LIPUS体外刺激hASCs细胞生长,采用Scepter 2.0手持细胞计数器进行细胞计数,运用荧光倒置显微镜观察细胞形态,运用流式细胞仪检测细胞表面标志物,并进行干细胞临床前质量检测.结果 刺激组50 mW/cm2和60 mW/cm2的LIPUS刺激强度均能促进细胞增殖,60 mW/cm2强度更显著,与对照组比较差异均具有统计学意义(P<0.05);细胞倍增时间比对照组缩短,差异具有统计学意义(P<0.05),刺激后刺激组细胞表面标志物无变化,增殖后干细胞质量检测符合标准要求.结论 LIPUS刺激可促进hASCs有效增殖,低强度脉冲场超声可作为干细胞体外扩增的新方法.

A new method for efficient amplification of human adipose-derived mesenchymal stem cells in vitro

Objective To provide a new method for efficient amplification of clinical stem cells in vitro us-ing low-intensity pulsed ultrasound (LIPUS)-stimulated proliferative human adipose-derived stem cells (hASCs). Methods hASCs cells growth was stimulated by LIPUS in vitro; cell counting was performed using a Scepter 2.0 handheld cell counter;cell morphology was observed by inverted fluorescence microscope;human cell surface marker was detected by flow cytometry and the preclinical quality of stem cells was measured. Results Cells in stimulation group treated with 50 mW/cm2 and 60 mW/cm2 LIPUS were both promoted in proliferation, but 60 mW/cm2 intensity was more effective (P<0.05), of which cell doubling time was significantly shorten compared to that in control group (P<0.05). There was no changes in cell surface markers of the stimulation group, and stem cell quality testing met the re-quirements of standard after proliferation. Conclusion LIPUS stimulation may promote effective proliferation of hASCs, and low-intensity pulsed ultrasound may be used as a new method for expansion of stem cells in vitro.