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评价人血小板裂解液与胎牛血清在间充质基质细胞培养中的应用。

Evaluation of human platelet lysate versus fetal bovine serum for culture of mesenchymal stromal cells.

机构信息

Helmholtz-Institute for Biomedical Engineering, RWTH Aachen University Medical School, Aachen, Germany.

Institute for Transfusion Medicine, University Hospital Essen, Essen, Germany.

出版信息

Cytotherapy. 2014 Feb;16(2):170-80. doi: 10.1016/j.jcyt.2013.11.004.

Abstract

Culture media for therapeutic cell preparations-such as mesenchymal stromal cells (MSCs)-usually comprise serum additives. Traditionally, fetal bovine serum is supplemented in basic research and in most clinical trials. Within the past years, many laboratories adapted their culture conditions to human platelet lysate (hPL), which further stimulates proliferation and expansion of MSCs. Particularly with regard to clinical application, human alternatives for fetal bovine serum are clearly to be preferred. hPL is generated from human platelet units by disruption of the platelet membrane, which is commonly performed by repeated freeze and thaw cycles. Such culture supplements are notoriously ill-defined, and many parameters contribute to batch-to-batch variation in hPL such as different amounts of plasma, a broad range of growth factors and donor-specific effects. The plasma components of hPL necessitate addition of anticoagulants such as heparins to prevent gelatinization of hPL medium, and their concentration must be standardized. Labels for description of hPL-such as "xenogen-free," "animal-free" and "serum free"-are not used consistently in the literature and may be misleading if not critically assessed. Further analysis of the precise composition of relevant growth factors, attachment factors, microRNAs and exosomes will pave the way for optimized and defined culture conditions. The use of hPL has several advantages and disadvantages: they must be taken into account because the choice of cell culture additive has major impact on cell preparations.

摘要

治疗性细胞制剂(如间充质基质细胞 [MSCs])的培养介质通常包含血清添加剂。在基础研究和大多数临床试验中,传统上补充胎牛血清。在过去的几年中,许多实验室已经将其培养条件适应于人血小板裂解物(hPL),这进一步刺激了 MSCs 的增殖和扩增。特别是在临床应用方面,显然应该优先选择人源替代胎牛血清。hPL 是通过破坏血小板膜从人血小板单位中产生的,这通常通过反复的冷冻和解冻循环来完成。这种培养补充剂的定义非常不明确,许多参数导致 hPL 的批次间变化,例如血浆量不同、生长因子范围广泛以及供体特异性效应。hPL 的血浆成分需要添加抗凝剂,如肝素,以防止 hPL 培养基的凝胶化,并且必须标准化其浓度。hPL 的标签描述,如“无异种”、“无动物”和“无血清”,在文献中没有一致使用,如果不进行严格评估,可能会产生误导。进一步分析相关生长因子、附着因子、microRNAs 和外泌体的精确组成将为优化和定义培养条件铺平道路。hPL 的使用有其优点和缺点:必须考虑到这一点,因为细胞培养添加剂的选择对细胞制剂有重大影响。

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