Fazzina R, Iudicone P, Mariotti A, Fioravanti D, Procoli A, Cicchetti E, Scambia G, Bonanno G, Pierelli L
InScientiaFides Foundation, San Marino, Republic of San Marino.
Immunohematology and Transfusion Medicine, San Camillo-Forlanini Hospital, Rome, Italy.
Cytotechnology. 2016 Aug;68(4):1185-95. doi: 10.1007/s10616-015-9878-5. Epub 2015 May 6.
Alternatives to the use of fetal bovine serum (FBS) have been investigated to ensure xeno-free growth condition. In this study we evaluated the efficacy of human platelet lysate (PL) as a substitute of FBS for the in vitro culture of some human cell lines. PL was obtained by pools of pathogen inactivated human donor platelet (PLT) concentrates. Human leukemia cell lines (KG-1, K562, JURKAT, HL-60) and epithelial tumor cell lines (HeLa and MCF-7) were cultured with either FBS or PL. Changes in cell proliferation, viability, morphology, surface markers and cell cycle were evaluated for each cell line. Functional characteristics were analysed by drug sensitivity test and cytotoxicity assay. Our results demonstrated that PL can support growth and expansion of all cell lines, although the cells cultured in presence of PL experienced a less massive proliferation compared to those grown with FBS. We found a comparable percentage of viable specific marker-expressing cells in both conditions, confirming lineage fidelity in all cultures. Functionality assays showed that cells in both FBS- and PL-supported cultures maintained their normal responsiveness to adriamycin and NK cell-mediated lysis. Our findings indicate that PL is a feasible serum substitute for supporting growth and propagation of haematopoietic and epithelial cell lines with many advantages from a perspective of process standardization, ethicality and product safety.
为确保无外源物生长条件，人们已对胎牛血清（FBS）的替代物展开研究。在本研究中，我们评估了人血小板裂解液（PL）作为FBS替代品用于某些人细胞系体外培养的效果。PL由经病原体灭活的人供体血小板（PLT）浓缩物混合制备而成。人白血病细胞系（KG-1、K562、JURKAT、HL-60）和上皮肿瘤细胞系（HeLa和MCF-7）分别用FBS或PL进行培养。对每个细胞系的细胞增殖、活力、形态、表面标志物和细胞周期的变化进行了评估。通过药物敏感性试验和细胞毒性测定分析功能特性。我们的结果表明，PL能够支持所有细胞系的生长和扩增，尽管与用FBS培养的细胞相比，在PL存在下培养的细胞增殖程度较低。我们发现在两种条件下表达特异性标志物的存活细胞百分比相当，证实了所有培养物中的谱系保真度。功能分析表明，在FBS和PL支持的培养物中的细胞对阿霉素和NK细胞介导的裂解均保持正常反应性。我们的研究结果表明，从工艺标准化、伦理和产品安全的角度来看，PL是一种可行的血清替代品，可用于支持造血和上皮细胞系的生长和增殖，具有诸多优势。
