Effect of activated autologous platelet-rich plasma on proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro.
Effect of activated autologous platelet-rich plasma on proliferation and osteogenic differentiation of human adipose-derived stem cells in vitro.
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DOI：
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发表时间：
2015-02
影响因子：
2.2
通讯作者：
Fang-tian Xu;Hong-Mian Li;Q. Yin;Zhijie Liang;Min-Hong Huang;Guang-Yi Chi;Lu Huang;Da-lie Liu;Hua Nan
中科院分区：
文献类型：
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作者：
Fang-tian Xu;Hong-Mian Li;Q. Yin;Zhijie Liang;Min-Hong Huang;Guang-Yi Chi;Lu Huang;Da-lie Liu;Hua Nan
目的：探讨活化的自体富血小板血浆(PRP)能否促进人脂肪干细胞(HASCs)的体外增殖和成骨分化。从脂肪抽提液中分离出hASCs，培养至第3代后，具有特异的细胞标志和多向分化能力。从同一患者的人外周血中收集并激活PRP。分别用正常成骨诱导培养液(A组)或成骨诱导培养液+5%、10%、20%、40%PRP(B、C、D、E组)处理体外培养的hASCs。CCK-8比色法检测细胞增殖。用实时定量聚合酶链式反应(QPCR)检测成骨标志基因碱性磷酸酶(ALP)、骨桥蛋白(OPN)、骨钙素(OCN)和核心结合因子α1(Cbfa1)的基因表达。结果显示，不同浓度的活化自体PRP与未加PRP组相比，在增殖期显著促进hASCs的生长，并呈现剂量-反应关系。在成骨诱导期的7d和14d时间点，PRP组的ALP活性随着PRP浓度的增加而逐渐升高，并呈剂量-反应关系。在成骨诱导期21d时间点，PRP组ALP、OPN、OCN和Cbfa1的矿化和mRNA相对表达均明显高于未加PRP组，且呈剂量-反应关系。本研究表明，不同浓度的活化自体PRP在体外能促进hASCs早期的增殖和后期的成骨分化。此外，激活的自体PRP对体外培养的hASCs的细胞增殖和成骨分化的影响呈剂量依赖性。
To investigate whether activated autologous platelet-rich plasma (PRP) can promote proliferation and osteogenic differentiation of human adipose-derived stem cells (hASCs) in vitro. hASCs were isolated from lipo-aspirates, and characterized by specific cell markers and multilineage differentiation capacity after culturing to the 3(rd) passage. PRP was collected and activated from human peripheral blood of the same patient. Cultured hASCs were treated with normal osteogenic inductive media alone (group A, control) or osteogenic inductive media plus 5%, 10%, 20%, 40%PRP (group B, C, D, E, respectively). Cell proliferation was assessed by CCK-8 assay. mRNA expression of osteogenic marker genes including alkaline phosphatase (ALP), osteopontin (OPN), osteocalcin (OCN) and core binding factor alpha 1 (Cbfa1) were determined by Real-Time Quantitative PCR Analysis (qPCR). Data revealed that different concentrations of activated autologous PRP significantly promoted hASCs growth in the proliferation phase compared to the without PRP group and resulted in a dose-response relationship. At 7-d and 14-d time point of the osteogenic induced stage, ALP activity in PRP groups gradually increased with the increasing of concentrations of PRP and showed that dose-response relationship. At 21-d time point of the osteogenic induced stage, PRP groups make much more mineralization and mRNA relative expression of ALP, OPN, OCN and Cbfa1 than that without PRP groups and show that dose-response relationship. This study indicated that different concentrations of activated autologous PRP can promote cell proliferation at earlier stage and promote osteogenic differentiation at later stage of hASCs in vitro. Moreover, it displayed a dose-dependent effect of activated autologous PRP on cell proliferation and osteogenic differentiation of hASCs in vitro.