Abstract：
SH-SY5Y is a human neuroblastoma cell line that can be differentiated into several neuronal phenotypes, depending on culture conditions. For this reason, this cell line has been widely used as an in vitro model of neurodegenerative conditions, such as Parkinson\'s disease (PD). However, most studies published to date used fetal bovine serum (FBS) as culture medium supplement for SH-SY5Y cell differentiation. We report on the testing of human platelet lysate (hPL) as a culture medium supplement to support SH-SY5Y cell culture. Both standard hPL and a fibrinogen-depleted hPL (FD-hPL) formulation, which does not require the addition of anticoagulants to culture media, promoted an increase in SH-SY5Y cell proliferation in comparison to FBS, without compromising metabolic activity. SH-SY5Y cells cultured in hPL or FD-hPL also displayed a higher number of neurite extensions and stained positive for MAP2 and synaptophysin, in the absence of differentiation stimuli; reducing hPL or FD-hPL concentration to 1% v/v did not affect cell proliferation or metabolic activity. Furthermore, following treatment with retinoic acid (RA) and further stimulation with brain-derived neurotrophic factor (BDNF) and nerve growth factor beta (NGF-β), the percentage of SH-SY5Y cells stained positive for dopaminergic neuronal differentiation markers (tyrosine hydroxylase [TH] and Dopamine Transporter [DAT]) was higher in hPL or FD-hPL than in FBS, and gene expression of dopaminergic markers TH, DAT, and DR2 was also detected. Overall, the data herein presented supports the use of hPL to differentiate SH-SY5Y cells into a neuronal phenotype with dopaminergic features, and the adoption of FD-hPL as a fully xenogeneic free alternative to FBS to support the use of SH-SY5Y cells as a neurodegeneration model.
摘要：
SH-SY5Y是一种人类神经母细胞瘤细胞系，可以分化为几种神经元表型，取决于培养条件。出于这个原因,该细胞系已被广泛用作神经退行性疾病的体外模型，如帕金森病（PD）。然而,迄今为止发表的大多数研究都使用胎牛血清（FBS）作为SH-SY5Y细胞分化的培养基补充剂。我们报告了人血小板裂解物（hPL）作为培养基补充剂的测试，以支持SH-SY5Y细胞培养。标准hPL和纤维蛋白原耗尽的hPL（FD-hPL）制剂，不需要在培养基中添加抗凝剂，与FBS相比，促进SH-SY5Y细胞增殖的增加，不损害代谢活动。在hPL或FD-hPL中培养的SH-SY5Y细胞也显示出更多的神经突延伸，并且MAP2和突触素染色呈阳性，在没有分化刺激的情况下;将hPL或FD-hPL浓度降低至1％v/v不影响细胞增殖或代谢活性。此外，在用维甲酸（RA）和脑源性神经营养因子（BDNF）和神经生长因子β（NGF-β）进一步刺激后，hPL或FD-hPL中多巴胺能神经元分化标志物（酪氨酸羟化酶[TH]和多巴胺转运蛋白[DAT]）染色阳性的SH-SY5Y细胞百分比高于FBS，多巴胺能标记物TH的基因表达，DAT,并且还检测到DR2。总的来说,本文提供的数据支持使用hPL将SH-SY5Y细胞分化为具有多巴胺能特征的神经元表型，以及采用FD-hPL作为FBS的完全异种游离替代品，以支持使用SH-SY5Y细胞作为神经变性模型。
