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骨髓间充质干细胞长期培养中神经营养因子和 TH、Nurr1、巢蛋白基因的自发表达。

Spontaneous Expression of Neurotrophic Factors and TH, Nurr1, Nestin Genes in Long-term Culture of Bone Marrow Mesenchymal Stem Cells.

机构信息

Department of Biology, Damghan University, Damghan, Iran.

出版信息

Cell J. 2012 Winter;13(4):243-50. Epub 2011 Dec 22.

Abstract

OBJECTIVE

It has been reported that rat bone marrow stromal cells (BMSCs) can be spontaneously differentiated into neural-like cells without any supplemental growth factors and/or chemical treatment after long-term culture.This study aims to determineWhether, growth factors secreted by MSCs could induce self-differentiation into neural-like cells in a long-term culture.

MATERIALS AND METHODS

THIS STUDY CONSISTED OF TWO GROUPS: i. rat BMSCs (passage 5) were cultured in alfa- minimal essential medium (α-MEM) and 10% fetal bovine serum (FBS) without the addition of inducer and exchanging medium for three weeks, as the experimental group and ii.rat BMSCs (passage 5) as the control group. Each group was analysed by reverse transcriptase polymerase chain reaction (RT-PCR) to evaluate the expressions of neurotrophic factors and neural marker genes. Statistical analyses were carried out using one-way analysis of variance (ANOVA) and Tukey's multiple comparison with SPSS software (version 16). P< 0.05 was considered statistically significant.

RESULTS

The experimental group (fifth passage of BMSCs) obtained from adult rats spontaneously differentiated into neural precursor cells after long-term culture. Cultured cells expressed tyrosine hydroxylase (TH), Nurr1 and nestin genes. Furthermore, some growing cells in suspension became neurosphere-like. Self-differentiated rat MSCs (SDrMSCs) expressed significantly higher levels of NGF (0.96 ± 0.16), nestin (0.63 ± 0.08), and Nurr1 (0.80 ± 0.10) genes (p<0.05).

CONCLUSION

In this study, we reported that rMSCs in long-term culture underwent spontaneous transformation to neural precursors without the supplement of growth factors and specific chemicals. Cells expressed neural markers such as: TH, Nurr1, and nestin genes.

摘要

目的

已有报道称,大鼠骨髓基质细胞(BMSCs)在长期培养后,无需添加任何补充生长因子和/或化学物质,即可自发分化为神经样细胞。本研究旨在确定MSC 分泌的生长因子是否能在长期培养中诱导其自我分化为神经样细胞。

材料与方法

本研究包括两组:i. 大鼠 BMSCs(第 5 代)在无诱导剂的α-最小必需培养基(α-MEM)和 10%胎牛血清(FBS)中培养,且不更换培养基,持续 3 周,作为实验组;ii. 大鼠 BMSCs(第 5 代)作为对照组。每组均通过逆转录聚合酶链反应(RT-PCR)分析,评估神经营养因子和神经标记基因的表达。采用 SPSS 软件(版本 16)的单因素方差分析(ANOVA)和 Tukey 多重比较进行统计学分析。P<0.05 被认为具有统计学意义。

结果

从成年大鼠中获得的实验组(第 5 代 BMSCs)在长期培养后自发分化为神经前体细胞。培养细胞表达酪氨酸羟化酶(TH)、Nurr1 和巢蛋白基因。此外,一些悬浮生长的细胞形成神经球样结构。自我分化的大鼠 MSC(SDrMSCs)表达的 NGF(0.96±0.16)、巢蛋白(0.63±0.08)和 Nurr1(0.80±0.10)基因水平显著更高(p<0.05)。

结论

在本研究中,我们报道了大鼠 MSC 在长期培养中无需补充生长因子和特定化学物质即可自发转化为神经前体细胞。细胞表达神经标记物,如:TH、Nurr1 和巢蛋白基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5f87/3584480/d93480b1a0fc/Cell-J-13-243-g01.jpg

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