Life and Health Science Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal.
Stem Cell Res Ther. 2012 May 2;3(3):18. doi: 10.1186/scrt109.
It is hypothesized that administration of stromal/stem cells isolated from the adipose tissue (ASCs) and umbilical cord (HUCPVCs) can ameliorate the injured central nervous system (CNS). It is still not clear, however, whether they have similar or opposite effects on primary cultures of neuronal populations. The objective of the present work was to determine if ASCs and HUCPVCs preferentially act, or not, on specific cell populations within the CNS.
Primary cultures of hippocampal neurons were exposed to ASCs and HUCPVCs conditioned media (CM) (obtained 24, 48, 72 and 96 hours after three days of culture) for one week.
Cell viability experiments (MTS (3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2(4-sulfophenyl)-2H tetrazolium) test) revealed that CM obtained from both cell populations at all time points did not cause any deleterious effects on neuronal cells. In fact, it was determined that whenever the ASCs CM were supplemented with basic fibroblast growth factor (bFGF) and B27, there was a significant increase in the metabolic viability and neuronal cell density of the cultures. On the other hand, in the absence of CM supplementation, it was the HUCPVCs secretome that had the highest impact on the metabolic viability and cell density. In an attempt to unveil which factors could be involved in the observed effects, a screening for the presence of bFGF, nerve growth factor (NGF), stem cell factor (SCF), hepatocyte growth factors (HGF) and vascular endothelial growth factor (VEGF) in the CM was performed. Results revealed the presence of all these factors in ASCs CM, except bFGF; in contrast, in HUCPVCs CM it was only possible to detect robust NGF expression.
Overall, the results confirm important differences on the secretome of ASCs and HUCPVCs, which lead to distinct effects on the metabolic viability and neuronal cell densities in primary cultures of hippocampal neurons; however, the factor(s) that promote the stronger effect of the HUCPVCs CM in neuronal survival is(are) still to be identified.
据推测，从脂肪组织（ASCs）和脐带（HUCPVCs）中分离的基质/干细胞的给药可以改善受损的中枢神经系统（CNS）。然而，目前尚不清楚它们对原代神经元群体培养物是否具有相似或相反的作用。本工作的目的是确定 ASCs 和 HUCPVCs 是否优先作用于 CNS 内的特定细胞群体。
将海马神经元原代培养物暴露于 ASC 和 HUCPVC 条件培养基（CM）（在培养 3 天后的 24、48、72 和 96 小时获得）中 1 周。
细胞活力实验（MTS（3-（4,5-二甲基噻唑-2-基）-5-（3-羧基甲氧基苯基）-2（4-磺基苯基）-2H 四唑）试验）表明，来自两种细胞群体的 CM 在所有时间点均未对神经元细胞造成任何有害影响。事实上，确定只要 ASC CM 补充碱性成纤维细胞生长因子（bFGF）和 B27，培养物的代谢活力和神经元细胞密度就会显著增加。另一方面，在没有 CM 补充的情况下，HUCPVCs 分泌组对代谢活力和细胞密度的影响最大。为了揭示可能参与观察到的效应的因素，对 CM 中 bFGF、神经生长因子（NGF）、干细胞因子（SCF）、肝细胞生长因子（HGF）和血管内皮生长因子（VEGF）的存在进行了筛选。结果表明，除了 bFGF 之外，所有这些因素都存在于 ASC CM 中；相反，在 HUCPVCs CM 中，只能检测到强 NGF 表达。
总的来说，结果证实了 ASCs 和 HUCPVCs 的分泌组存在重要差异，这导致对海马神经元原代培养物的代谢活力和神经元细胞密度产生不同的影响；然而，促进 HUCPVCs CM 对神经元存活的更强作用的因素（s）仍有待确定。
