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富含血小板血浆和血小板贫化血浆可能在癌细胞培养中发挥支持作用:替代胎牛血清?

Platelet-Rich and Platelet-Poor Plasma Might Play Supportive Roles in Cancer Cell Culture: A Replacement for Fetal Bovine Serum?

机构信息

Stem Cell Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Department of Biochemistry and Molecular Biology, LSUHSC, School of Medicine, New Orleans, LA, United States.

出版信息

Anticancer Agents Med Chem. 2021 Oct 28;21(16):2236-2242. doi: 10.2174/1871520621999210101225912.

Abstract

BACKGROUND

Platelet-Rich (PRP) and Platelet-Poor plasma (PPP) are widely used in research and clinical platforms mainly due to their capacities to enhance cell growth. Although the short half-life (5 days) and the high price of platelet products pose challenges regarding their usage, they maintain the growth regulatory functions for weeks. Thus, we aimed to assess the supplementary values of these products in human CCRF- CEM cancer cells. Mechanistically, we also checked if the PRP/PPP treatment enhances YKL-40 expression as a known protein regulating cell growth.

METHODS

The PRP/PPP was prepared from healthy donors using manual stepwise centrifugation and phase separation. The viability of the cells treated with gradient PRP/PPP concentrations (2, 5, 10, and 15%) was measured by the MTT assay. The YKL-40 mRNA and protein levels were assessed using qRT-PCR and western blotting. The data were compared to FBS-treated cells.

RESULTS

Our findings revealed that the cells treated by PRP/PPP not only were morphologically comparable to those treated by FBS but also showed greater viability at the concentrations of 10 and 15%. Moreover, it was shown that PRP/PPP induce cell culture support, at least in part, via inducing YKL-40 expression at both mRNA and protein levels in a time- and dose-dependent manner.

CONCLUSION

Collectively, by showing cell culture support comparable to FBS, the PRP/PPP might be used as good candidates to supplement the cancer cell culture and overcome concerns regarding the use of FBS as a non-human source in human cancer research.

摘要

背景

富含血小板(PRP)和贫血小板血浆(PPP)由于能够促进细胞生长而广泛应用于研究和临床平台。尽管血小板产品半衰期短(5 天)且价格昂贵,但其对细胞生长的调控功能可维持数周。因此,我们旨在评估这些产品在人 CCRF-CEM 癌细胞中的补充价值。从机制上讲,我们还检查了 PRP/PPP 处理是否会增强 YKL-40 表达,因为 YKL-40 是一种已知的调节细胞生长的蛋白。

方法

使用手动逐步离心和相分离法从健康供体中制备 PRP/PPP。通过 MTT 测定法测量用梯度 PRP/PPP 浓度(2、5、10 和 15%)处理的细胞的活力。使用 qRT-PCR 和 Western blot 评估 YKL-40 mRNA 和蛋白水平。将数据与 FBS 处理的细胞进行比较。

结果

我们的研究结果表明,PRP/PPP 处理的细胞不仅在形态上与 FBS 处理的细胞相似,而且在浓度为 10%和 15%时显示出更高的活力。此外,研究表明 PRP/PPP 通过在时间和剂量依赖性方式诱导 YKL-40 在 mRNA 和蛋白水平上的表达来诱导细胞培养支持,至少部分如此。

结论

总之,通过显示与 FBS 相当的细胞培养支持,PRP/PPP 可作为补充癌细胞培养的良好候选物,并克服使用 FBS 作为非人类来源在人类癌症研究中的担忧。

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