Platelet-derived growth factor (PDGF)-AA/AB in human serum are potential indicators of the proliferative capacity of human synovial mesenchymal stem cells.
Platelet-derived growth factor (PDGF)-AA/AB in human serum are potential indicators of the proliferative capacity of human synovial mesenchymal stem cells.
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DOI：
10.1186/s13287-015-0239-2
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发表时间：
2015-12-10
影响因子：
7.5
通讯作者：
Sekiya I
中科院分区：
文献类型：
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作者：
Mizuno M;Katano H;Otabe K;Komori K;Matsumoto Y;Fujii S;Ozeki N;Tsuji K;Koga H;Muneta T;Matsuyama A;Sekiya I
对于人骨髓间充质干细胞（MSCs）的扩增，在临床情况下，自体人血清比胎牛血清更安全。使用自体人血清的问题之一是其对MSC的增殖作用在供体之间变化很大。本研究的三重目标是：（1）证明制备人血清的改进方法;（2）鉴定预测增殖潜力的生长因子;和（3）鉴定促进人血清中MSC增殖的细胞因子。使用含有玻璃珠的封闭袋系统收集新鲜血液。将袋在20 °C下摇动30分钟以进行快速制备，或在4 °C下保持静止24小时以进行缓慢制备。将来自四个供体的第0代滑膜MSC与来自四个不同供体的10%常规快速制备血清或改良的缓慢制备血清一起培养。为了进行集落形成单位测定，在这些血清中培养滑膜MSC。用细胞因子芯片分析血清蛋白表达谱。根据克隆形成能力与蛋白表达量的相关性推测候选蛋白。作为候选蛋白的评估，检查滑膜MSC的增殖能力、表面标记表型和分化能力。与快速制备血清相比，慢速制备血清的菌落总数显著增加，其中促血管生成素-1、BDNF、EGF、ENA-78、IGFBP-2、血小板源性生长因子（PDGF）-AA、PDGF-AB/BB、RANTES和TfR等9种蛋白的表达量增加了2倍。集落数与PDGF-AA/AB浓度呈正相关。外源性PDGF-AA能显著促进滑膜间充质干细胞的增殖，而PDGFR抑制剂则抑制增殖，PDGFs和PDGFR抑制剂对滑膜间充质干细胞表面抗原表位无影响。PDGFs或PDGFR抑制剂预处理不影响滑膜间充质干细胞的软骨形成、脂肪形成或钙化潜能。慢制备血清含有较高浓度的PDGF-AA/AB，滑膜间充质干细胞的殖民地形成数增加。PDGF-AA/AB是人血清增殖潜能的指标。外源性PDGF-AA增加滑膜间充质干细胞的增殖，而不改变表面表位和分化潜能。
For expansion of human mesenchymal stem cells (MSCs), autologous human serum is safer than fetal bovine serum in clinical situations. One of the problems with the use of autologous human serum is that its proliferative effect on MSCs varies widely between donors. The threefold goals of this study were: (1) to demonstrate an improved method for preparing human serum; (2) to identify growth factors predictive of proliferative potential; and (3) to identify a cytokine to promote MSC proliferation in human serum. Fresh blood was collected using a closed bag system containing glass beads. The bag was shaken at 20 °C for 30 minutes for rapid preparation, or kept stationary at 4 °C for 24 hours for slow preparation. Passage 0 synovial MSCs derived from four donors were cultured with 10 % conventional rapid preparation serum or modified slow preparation serum from four different donors. To perform the colony-forming unit assay, synovial MSCs were cultured in these serums. The protein expression profile in serum was analyzed using cytokine array. The candidate proteins were speculated from the correlation between the colony-forming ability and protein expression. As an evaluation of the candidate proteins, proliferation ability, surface marker phenotype and differentiation capability of synovial MSCs were examined. Compared with rapid preparation serum, slow preparation serum resulted in a significantly higher total colony number and twofold higher expression levels of nine proteins (angiopoietin-1, BDNF, EGF, ENA-78, IGFBP-2, platelet-derived growth factor (PDGF)-AA, PDGF-AB/BB, RANTES and TfR). Colony number was positively correlated with PDGF-AA/AB concentrations. Exogenous PDGF-AA significantly promoted proliferation of synovial MSCs, whereas PDGF receptor (PDGFR) inhibitor decreased it. Addition of PDGFs or PDGFR inhibitor did not affect surface epitopes of synovial MSCs. Pretreatment with PDGFs or PDGFR inhibitor did not affect chondrogenic, adipogenic, or calcification potentials of synovial MSCs. Slow preparation serum contained higher concentrations of PDGF-AA/AB and increased the colony formation number of synovial MSCs. PDGF-AA/AB were indicators of the proliferative potential of human serum. Exogenous PDGF-AA increased proliferation of synovial MSCs without alteration of surface epitopes and differentiation potentials.