Epidermal growth factor, basic fibroblast growth factor and platelet-derived growth factor-bb can substitute for fetal bovine serum and compete with human platelet-rich plasma in the ex vivo expansion of mesenchymal stromal cells derived from adipose tissue
Epidermal growth factor, basic fibroblast growth factor and platelet-derived growth factor-bb can substitute for fetal bovine serum and compete with human platelet-rich plasma in the ex vivo expansion of mesenchymal stromal cells derived from adipose tissue
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DOI：
10.3109/14653249.2011.583232
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发表时间：
2011-09-01
期刊：
影响因子：
4.5
通讯作者：
Rodeghiero, Francesco
中科院分区：
文献类型：
--
作者：
Chieregato, Katia;Castegnaro, Silvia;Rodeghiero, Francesco
背景目标。人间充质基质细胞（MSC）是具有自我更新能力、长期存活能力和多向分化潜能的多能细胞。为了避免异种血清的使用，我们分析了四种不同的培养基补充剂对脂肪组织来源的MSC（ADSC）扩增和分化的影响。方法.我们比较了胎牛血清（FBS）与10%人富血小板血浆（hPRP），3%人贫血小板血浆（hPPP）和细胞因子鸡尾酒组成的表皮生长因子（EGF），碱性成纤维细胞生长因子（bFGF）和血小板衍生生长因子-BB（PDGFbb）添加到3%的hPPP。该混合物是通过测试单独添加或与hPPP组合添加的EGF、bFGF、粒细胞集落刺激因子（G-CSF）、肝细胞生长因子（HGF）、胰岛素样生长因子（IGF-I）、PDGFbb和转化生长因子（TGF）-β 1而开发的。结果我们的数据表明，在补充有hPPP的培养基中加入EGF、bFGF和PDGFbb，可从150-200 mL自体全血获得，比FBS更好地支持ADSC扩增，如通过累积群体倍增（cPD; 15.0 +/-0.5对9.4 +/-2.8）所证实的。添加人富血小板血浆（hPRP）进一步改善了ADSC增殖（cPD 20.0 +/- 1.2），但hPRP的实现存在一个主要缺点，需要1000-1200 mL自体或供体全血。培养基补充剂不影响ADSC表型：它们表达缺乏造血抗原的CD 105、CD 90和CD 44。暴露于所提出的混合物或hPRP增加了成脂和成骨分化。结论.在hPPP中加入EGF、bFGF和PDGFbb可以确保足够数量的ADSC用于临床应用，避免使用动物血清，代表再生医学的新方法。
Background aims. Human mesenchymal stromal cells (MSC) are multipotent cells possessing self-renewal capacity, long-term viability and multilineage potential. We analyzed the effect of four different medium supplements on the expansion and differentiation of adipose tissue-derived MSC (ADSC) in order to avoid the use of xenogeneic serum. Methods. We compared fetal bovine serum (FBS) with 10% human platelet-rich plasma (hPRP), 3% human platelet-poor plasma (hPPP) and with a cytokine cocktail composed of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and platelet-derived growth factor-bb (PDGFbb) added to 3% hPPP. This mixture was developed testing EGF, bFGF, granulocyte-colony-stimulating factor (G-CSF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF-I), PDGFbb and transforming growth factor (TGF)-beta 1 added alone or in combination with hPPP. Results. Our data demonstrate that the addition of EGF, bFGF and PDGFbb, in a medium supplemented with hPPP, obtainable from 150-200 mL whole autologous blood, supports ADSC expansion better than FBS, as confirmed by cumulative population doublings (cPD; 15.0 +/- 0.5 versus 9.4 +/- 2.8). The addition of human platelet-rich plasma (hPRP) further improved ADSC proliferation (cPD 20.0 +/- 1.2), but the achievement of hPRP presented a major drawback, requiring 1000-1200 mL autologous or donor whole blood. The medium supplements did not influence ADSC phenotype: they expressed CD105, CD90 and CD44 lacking hematopoietic antigens. The exposure to the proposed cocktail or to hPRP increased adipogenic and osteogenic differentiation. Conclusions. The addition of EGF, bFGF and PDGFbb to hPPP could ensure a sufficient number of ADSC for clinical applications, avoiding the use of animal serum and representing a novel approach in regenerative medicine.