Evaluation of the viability and osteogenic differentiation of cryopreserved human adipose-derived stem cells
Evaluation of the viability and osteogenic differentiation of cryopreserved human adipose-derived stem cells
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DOI：
10.1016/j.cryobiol.2008.04.002
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发表时间：
2008-08-01
期刊：
影响因子：
2.7
通讯作者：
Cao, Yilin
中科院分区：
文献类型：
--
作者：
Liu, Guangpeng;Zhou, Heng;Cao, Yilin
人脂肪干细胞具有向成骨细胞分化的能力，在组织工程骨的构建中具有潜在的应用价值，因此需要一种可靠的细胞储存方法。本研究的目的是确定是否有一个简单的方法，冷冻保存与10%的Me 2SO作为保护剂的增殖潜力和成骨分化的ASCs从新鲜的人脂肪组织分离的影响。从6个人脂肪抽吸物中收获ASC，并将每个减半用于在液氮中冷冻保存2周或用于对照培养。将来自第二代的细胞以5000个细胞/孔的密度接种在24孔板中，并在有或没有成骨培养基的情况下培养14天。用流式细胞仪检测细胞表面抗原。使用细胞DNA测定评价两个群体的增殖率。为了检测冻存和非冻存人群的成骨分化，应用成骨细胞蛋白产生（碱性磷酸酶和骨钙素）和细胞外基质钙化（钙含量）的测定。成骨细胞相关基因的表达也进行了分析，使用逆转录聚合酶链反应。这些结果表明，冷冻保存对人ASCs的表型、增殖和成骨分化没有影响，表明冷冻保存的人ASCs可能用于骨组织工程。(C)2008年爱思唯尔公司All rights reserved.
Human adipose-derived stem cells (ASCs) have the ability to differentiate into osteoblasts and thus the potential therapeutic use to tissue-engineer bone, so a reliable method for cell storage is necessary. The aim of this study was to determine whether a simple method of cryopreservation with 10% Me2SO as a protectant had an effect on proliferation potential and osteogenic differentiation of ASCs isolated from fresh human adipose tissue. ASCs were harvested from 6 human lipoaspirates and each was halved for either cryopreservation in liquid nitrogen for 2 weeks or for control culture. Cells from the second-passage were plated at a density of 5000 cells/well in 24-well plates and cultured with or without osteogenic media for 14 days. Cell surface antigens were used to identify the cryopreserved ASCs by flow cytometry. The proliferation rate of both populations was evaluated using a cell DNA assay. To detect osteogenic differentiation of both the cryopreserved and non-cryopreserved populations, determination of osteoblastic protein production (alkaline phosphatase and osteocalcin) and excellular matrix calcification (calcium content) was applied. The expression of osteoblastic-associated genes was also analyzed using reverse-transcription polymerase chain reaction. These results demonstrate that cryopreservation has no effect on the phenotype, proliferation or osteogenic differentiation of human ASCs, showing cryopreserved human ASCs might be applied for bone tissue engineering. (C) 2008 Elsevier Inc. All rights reserved.