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富血小板血浆和胎牛血清对体外扩增骨髓来源人间充质基质细胞的影响差异。

Differential effect of platelet-rich plasma and fetal calf serum on bone marrow-derived human mesenchymal stromal cells expanded in vitro.

机构信息

University Hospital Carl Gustav Carus, Department of Haematology, Dresden, Germany.

出版信息

J Tissue Eng Regen Med. 2011 Aug;5(8):648-54. doi: 10.1002/term.359. Epub 2011 Jan 14.

DOI:10.1002/term.359
PMID:21774088
Abstract

Mesenchymal stromal cells (MSCs) derived from various sources have great potential for use in cell-based therapies. Since the proportion of primary MSCs contained in bone marrow or adipose tissue is low, plastic adherence and in vitro expansion are necessary to expand MSCs prior to clinical application. Human platelet-rich plasma has been introduced as an alternative serum source but functional differences have so far not been described. Here we cultured MSCs derived from human bone marrow in medium supplemented with either 10% fetal calf serum (FCS) or 5% and 10% platelet-rich plasma (PRP) until the first or second passage. Parameters under investigation were cell yield, clonogenicity, phenotype as well as migratory and differentiation potential. In addition, the secretion of SDF-1α and the induced migration of CD34(+) haematopoietic stem cells (HSCs) were investigated with regard to the different serum source. The use of PRP resulted in a significantly higher expansion rate and yield at passages 0 and 1. In addition, the level of secreted SDF-1α was significantly increased in the supernatant of MSCs cultured with FCS instead of human PRP. Consistent with this, the migration capacity of MSCs cultured with 10% FCS as well as their capability to induce the migration of CD34(+) haematopoietic progenitors in a transwell assay was higher. Our results demonstrate that human PRP can be seen as an alternative serum source to FCS for MSC cultivation. However, the requirements of the specific clinical application must be carefully considered before the respective serum source is selected.

摘要

间充质基质细胞(MSCs)来源于各种来源,在基于细胞的治疗中有很大的应用潜力。由于骨髓或脂肪组织中所含的原代 MSCs 比例较低,因此在临床应用前需要通过塑料贴附和体外扩增来扩增 MSCs。已经引入了富含血小板的血浆作为替代血清来源,但迄今为止尚未描述其功能差异。在这里,我们在补充有 10%胎牛血清(FCS)或 5%和 10%富含血小板的血浆(PRP)的培养基中培养来源于人骨髓的 MSCs,直到第 1 或第 2 代。研究的参数包括细胞产量、集落形成能力、表型以及迁移和分化潜能。此外,还研究了不同血清来源对 SDF-1α分泌和 CD34+造血干细胞(HSCs)诱导迁移的影响。使用 PRP 可在第 0 代和第 1 代时显著提高扩增率和产量。此外,与使用人 PRP 相比,在 FCS 培养的 MSC 的上清液中 SDF-1α 的分泌水平显著增加。与此一致的是,用 10% FCS 培养的 MSC 的迁移能力以及它们在 Transwell 测定中诱导 CD34+造血祖细胞迁移的能力更高。我们的结果表明,富含血小板的血浆可以作为 MSC 培养的替代血清来源。然而,在选择各自的血清来源之前,必须仔细考虑特定临床应用的要求。

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