人血小板裂解物增强沃顿果冻间充质干细胞的增殖

Andreas Ardhika Antoninus , Wahyu Widowati , Laura Wijaya , Dwi Agustina , Sugiarto Puradisastra , Sutiman B. Sumitro , M.Aris Widodo , Indra Bachtiar
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引用次数: 23

摘要

本研究旨在阐明人血小板裂解物(huPL)作为培养间充质干细胞(MSC)的无异种培养基的替代品的潜力。在这项工作中,分离、表征了沃顿果冻衍生的MSCs(WJ MSCs;n=5),并在用AB型贫血小板血浆(huPL–ABO)重建的O型血小板衍生的huPL中培养。在两种不同条件下培养来自五个供体的WJ MSCs[补充有20%胎牛血清(FBS)和0.250 mg/mL、0.125 mg/mL、0.063 mg/mL和0.031 mg/mL huPL的细胞培养物]。评估了生长动力学、细胞表面标志物和向脂肪、软骨和成骨谱系的体外分化潜力。我们的结果表明,与在FBS中培养的细胞相比,在huPL–ABO存在下培养的WJ MSCs显示出典型的成纤维细胞样形态,并且具有显著更高的细胞计数(p<;0.05)。此外,免疫表型分析的结果显示,在两种培养条件下,MSCs的特征相似。此外,间充质干细胞向脂肪、软骨和成骨谱系的分化没有观察到显著差异。培养基中huPL–ABO的最佳浓度为0.250 mg/mL和0.125 mg/mL。由于WJ MSCs群体倍增时间方面的浓度差异不显著,因此在FBS和huPL–ABO之间的所有比较中均使用0.125 mg/mL的浓度。huPL–ABO中胰岛素样生长因子-1、血小板衍生生长因子AB、血管内皮生长因子和转化生长因子-β1的平均浓度分别为287.89、47096.63、150.93和74817.76。与FBS培养条件相比,该方法产生的huPL–ABO能够不断提高WJ MSCs的增殖率,并缩短达到融合所需的时间。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Human platelet lysate enhances the proliferation of Wharton's jelly-derived mesenchymal stem cells

This study was performed to elucidate the potential of human platelet lysate (huPL) as an alternative to xeno-free media for culturing mesenchymal stem cells (MSCs). In this work, Wharton's jelly-derived MSCs (WJ-MSCs; n = 5) were isolated, characterized, and cultured in huPL derived from type O platelets reconstituted with type AB platelet-poor plasma (huPL–ABO). WJ-MSCs from five donors were cultured under two different conditions [cell culture supplemented with 20% fetal bovine serum (FBS) and 0.250 mg/mL, 0.125 mg/mL, 0.063 mg/mL, and 0.031 mg/mL huPL]. Growth kinetics, cell surface markers, and in vitro differentiation potential toward the adipogenic, chondrogenic, and osteogenic lineages were evaluated. Our results indicate that WJ-MSCs cultured in the presence of huPL–ABO showed typical fibroblast-like morphology and had a significantly higher cell count (p < 0.05), compared with those cultured in FBS. Furthermore, results of immunophenotyping assay showed similar MSCs characteristics for both culture conditions. In addition, no significant differences were observed in the MSCs differentiation toward the adipogenic, chondrogenic, and osteogenic lineages. The optimal concentrations of huPL–ABO in the culture medium were 0.250 mg/mL and 0.125 mg/mL. Because of insignificant differences between the concentrations in terms of WJ-MSCs population doubling time, the concentration of 0.125 mg/mL was used in all comparisons between FBS and huPL–ABO. The average concentrations of insulin-like growth factor-1, platelet-derived growth factor-AB, vascular endothelial growth factor, and transforming growth factor-β1 in huPL–ABO were 287.89 pg/mg, 47,096.63 pg/mg, 150.93 pg/mg, and 74,817.76 pg/mg, respectively. huPL–ABO produced by this method was able to enhance the WJ-MSCs proliferation rate constantly and decrease the required time to reach confluence compared with the FBS culture condition.

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