人富血小板血浆补充培养基中间充质基质细胞的增殖、基因表达和蛋白质生成

Mesenchymal stromal cell proliferation, gene expression and protein production in human platelet-rich plasma-supplemented media.

作者信息

Amable Paola Romina, Teixeira Marcus Vinicius Telles, Carias Rosana Bizon Vieira, Granjeiro José Mauro, Borojevic Radovan

机构信息

Excellion Biomedical Services S.A., Petrópolis, Rio de Janeiro, Brazil.

Excellion Biomedical Services S.A., Petrópolis, Rio de Janeiro, Brazil; National Institute of Metrology, Quality and Technology, Xerém, Rio de Janeiro, Brazil.

出版信息

PLoS One. 2014 Aug 12;9(8):e104662. doi: 10.1371/journal.pone.0104662. eCollection 2014.

DOI:10.1371/journal.pone.0104662

PMID:25115920

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4130592/

Abstract

BACKGROUND

Platelet-rich plasma (PRP) is increasingly used as a cell culture supplement, in order to reduce the contact of human cells with animal-derived products during in vitro expansion. The effect of supplementation changes on cell growth and protein production is not fully characterized.

METHODS

Human mesenchymal stromal cells from bone marrow, adipose tissue and Wharton's Jelly were isolated and cultured in PRP-supplemented media. Proliferation, in vitro differentiation, expression of cell surface markers, mRNA expression of key genes and protein secretion were quantified.

RESULTS

10% PRP sustained five to tenfold increased cell proliferation as compared to 10% fetal bovine serum. Regarding cell differentiation, PRP reduced adipogenic differentiation and increased calcium deposits in bone marrow and adipose tissue-mesenchymal stromal cells. Wharton's Jelly derived mesenchymal stromal cells secreted higher concentrations of chemokines and growth factors than other mesenchymal stromal cells when cultured in PRP-supplemented media. Bone marrow derived mesenchymal stromal cells secreted higher concentrations of pro-inflammatory and pro-angiogenic proteins. Mesenchymal stromal cells isolated from adipose tissue secreted higher amounts of extracellular matrix components.

CONCLUSIONS

Mesenchymal stromal cells purified from different tissues have distinct properties regarding differentiation, angiogenic, inflammatory and matrix remodeling potential when cultured in PRP supplemented media. These abilities should be further characterized in order to choose the best protocols for their therapeutic use.

摘要

背景

富血小板血浆（PRP）越来越多地用作细胞培养补充剂，以减少体外扩增过程中人类细胞与动物源产品的接触。补充物变化对细胞生长和蛋白质产生的影响尚未完全明确。

方法

从骨髓、脂肪组织和脐带华通氏胶中分离人骨髓间充质基质细胞，并在补充PRP的培养基中培养。对细胞增殖、体外分化、细胞表面标志物表达、关键基因的mRNA表达和蛋白质分泌进行定量分析。

结果

与10%胎牛血清相比，10%PRP使细胞增殖增加了五到十倍。关于细胞分化，PRP减少了骨髓和脂肪组织间充质基质细胞的成脂分化并增加了钙沉积。当在补充PRP的培养基中培养时，脐带华通氏胶来源的间充质基质细胞分泌的趋化因子和生长因子浓度高于其他间充质基质细胞。骨髓来源的间充质基质细胞分泌更高浓度的促炎和促血管生成蛋白。从脂肪组织分离的间充质基质细胞分泌更多的细胞外基质成分。