低温制剂保存对脂肪间充质干细胞生物学特性的影响

发布时间：2019-02-11 17:04

【摘要】：背景:脂肪干细胞在GMP洁净车间制备完成后需要做成制剂才能给患者输注。低温制剂相比于冻存制剂有很多优势,然而关于低温短期保存对干细胞生物学特性影响的研究较少。目的:评价用含5%人血白蛋白的复方电解质低温保存24 h对脂肪间充质干细胞生物学特性的影响。方法:取第3-6代人脂肪间充质干细胞,以5×109 L-1的细胞浓度重悬在含5%人血白蛋白的复方电解质注射液中,将细胞悬液转移至冻存管中,置于冷链运输箱,2-8℃低温保存24 h。观察低温保存前后的细胞形态、贴壁能力、细胞活率、细胞直径及细胞免疫表型变化。结果与结论:(1)脂肪间充质干细胞低温保存24 h后,死细胞增多,细胞活率显著下降,但细胞活率仍大于80%,活细胞直径显著增大;(2)脂肪间充质干细胞低温保存24 h后,少量细胞为圆形,失去贴壁能力,大部分细胞能贴壁生长,形态为梭形,与细胞保存前贴壁形态相似;(3)脂肪间充质干细胞低温保存24 h后,HLA-DR、CD34和CD45均为阴性表达,阳性率低于2%;CD29、CD73和CD105均为阳性表达,阳性率高于95%,但保存后的细胞显著地由一群分为两群,部分细胞体积增大,向右移动;(4)结果表明,低温制剂保存未明显影响脂肪间充质干细胞的贴壁生长能力、存活率及免疫表型等干性生物学特性。
[Abstract]:Background: adipose stem cells need to be prepared after preparation in GMP clean workshop to be infused into patients. Cryopreservation has many advantages over cryopreservation, but there are few studies on the effects of cryopreservation on the biological characteristics of stem cells. Aim: to evaluate the effects of compound electrolyte containing 5% human serum albumin (HSA) on the biological characteristics of adipose mesenchymal stem cells (AMSCs) after cryopreservation for 24 h. Methods: human adipose mesenchymal stem cells (HMSCs) of passage 3-6 were resuspended in 5 脳 109 L-1 cell concentration in compound electrolyte injection containing 5% human serum albumin. The suspension was transferred to the cryopreserved tube and placed in a cold chain transport box. Stored at 2-8 鈩，

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