Dipartimento di Medicina Sperimentale and Centro di Eccellenza per la Ricerca Biomedica, Università degli Studi di Genova, Genova, Italy.
Eur J Immunol. 2011 Nov;41(11):3281-90. doi: 10.1002/eji.201141542. Epub 2011 Oct 18.
The use of fetal bovine serum (FBS) for the culture and expansion of mesenchymal stromal cells (MSCs) limits their possible clinical applications. Although some recent studies recommended substituting FBS with human platelet lysate (HPL) for the expansion of MSCs for clinical use, the functional capacity of the expanded cells has only been partially explored. 10% FBS and two other commercial FBS-containing media (MesenCult and MesenPro) were compared with 10% HPL-containing medium for their ability to support MSCs expansion and immunomodulation. We demonstrate that HPL sustained MSC proliferation and expansion in vitro. However, the cumulative cell numbers recovered were comparable with those obtained in MesenPro medium. Moreover, we show that HPL alters the expression of some relevant MSC surface molecules, namely the DNAM-1 ligands PVR and Nectin-2, the NKG2D ligand ULBP3, the adhesion molecules CD49d and αvβ3 and the fibroblast-associated protein. In addition, MSCs cultured in HPL displayed impaired inhibitory capacity on T-cell proliferation to alloantigen and NK-cell proliferation and cytotoxicity. Finally, they showed decreased constitutive PGE2 production while IL-6, IL-8 and RANTES secretion were upregulated. These results imply some limitations in the use of HPL for the expansion of MSCs to be used as immunomodulators in clinical applications.
使用胎牛血清(FBS)培养和扩增间充质基质细胞(MSCs)会限制其可能的临床应用。尽管最近的一些研究建议用人血小板裂解物(HPL)替代 FBS 来扩增 MSCs 以用于临床应用,但扩增细胞的功能能力仅得到了部分探索。本研究比较了含 10%FBS 的培养基和两种其他商业化含 FBS 的培养基(MesenCult 和 MesenPro)与含 10%HPL 的培养基,以评估它们支持 MSCs 扩增和免疫调节的能力。我们证明 HPL 能够支持 MSC 的体外增殖和扩增。然而,回收的细胞总数与 MesenPro 培养基中获得的数量相当。此外,我们还表明 HPL 改变了一些相关 MSC 表面分子的表达,即 DNAM-1 配体 PVR 和 Nectin-2、NKG2D 配体 ULBP3、黏附分子 CD49d 和 αvβ3 以及成纤维细胞相关蛋白。此外,在 HPL 中培养的 MSC 对同种异体抗原刺激的 T 细胞增殖和 NK 细胞增殖及细胞毒性的抑制能力受损。最后,它们表现出降低的基础 PGE2 产生,同时 IL-6、IL-8 和 RANTES 的分泌上调。这些结果表明,在将 HPL 用于扩增 MSCs 以用作临床应用中的免疫调节剂方面存在一些局限性。