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人脐带华通氏胶源性间充质干细胞向神经样细胞的分化

Human umbilical cord Wharton's Jelly-derived mesenchymal stem cells differentiation into nerve-like cells.

作者信息

Ma Lian, Feng Xue-yong, Cui Bing-lin, Law Frieda, Jiang Xue-wu, Yang Li-Ye, Xie Qing-dong, Huang Tian-hua

机构信息

Department of Pediatrics, Second Affiliated Hospital of Shantou University Medical College, Shantou 515041, China.

出版信息

Chin Med J (Engl). 2005 Dec 5;118(23):1987-93.

Abstract

BACKGROUND

The two most basic properties of mesenchymal stem cells (MSCs) are the capacities to self-renew indefinitely and differentiate into multiple cells and tissue types. The cells from human umbilical cord Wharton's Jelly have properties of MSCs and represent a rich source of primitive cells. This study was conducted to explore the possibility of inducing human umbilical cord Wharton's Jelly-derived MSCs to differentiate into nerve-like cells.

METHODS

MSCs were cultured from the Wharton's Jelly taken from human umbilical cord of babies delivered after full-term normal labor. Salvia miltiorrhiza and beta-mercaptoethanol were used to induce the human umbilical cord-derived MSCs to differentiate. The expression of neural protein markers was shown by immunocytochemistry. The induction process was monitored by phase contrast microscopy, electron microscopy (EM), and laser scanning confocal microscopy (LSCM). The pleiotrophin and nestin genes were measured by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTS

MSCs in the Wharton's Jelly were easily attainable and could be maintained and expanded in culture. They were positive for markers of MSCs, but negative for markers of hematopoietic cells and graft-versus-host disease (GVHD)-related cells. Treatment with Salvia miltiorrhiza caused Wharton's Jelly cells to undergo profound morphological changes. The induced MSCs developed rounded cell bodies with multiple neurite-like extensions. Eventually they developed processes that formed networks reminiscent of primary cultures of neurons. Salvia miltiorrhiza and beta-mercaptoethanol also induced MSCs to express nestin, beta-tubulinIII, neurofilament (NF) and glial fibrillary acidic protein (GFAP). It was confirmed by RT-PCR that MSCs could express pleiotrophin both before and after induction by Salvia miltiorrhiza. The expression was markedly enhanced after induction and the nestin gene was also expressed.

CONCLUSIONS

MSCs could be isolated from human umbilical cord Wharton's Jelly. They were capable of differentiating into nerve-like cells using Salvia miltiorrhiza or beta-mercaptoethanol. The induced MSCs not only underwent morphologic changes, but also expressed the neuron-related genes and neuronal cell markers. They may represent an alternative source of stem cells for central nervous system cell transplantation.

摘要

背景

间充质干细胞(MSCs)的两个最基本特性是无限自我更新以及分化为多种细胞和组织类型的能力。人脐带华通氏胶中的细胞具有间充质干细胞的特性,是原始细胞的丰富来源。本研究旨在探索诱导人脐带华通氏胶来源的间充质干细胞分化为神经样细胞的可能性。

方法

从足月正常分娩后婴儿的脐带中获取华通氏胶来培养间充质干细胞。使用丹参和β-巯基乙醇诱导人脐带来源的间充质干细胞分化。通过免疫细胞化学显示神经蛋白标志物的表达。通过相差显微镜、电子显微镜(EM)和激光扫描共聚焦显微镜(LSCM)监测诱导过程。通过逆转录-聚合酶链反应(RT-PCR)检测多效生长因子和巢蛋白基因。

结果

华通氏胶中的间充质干细胞易于获取,并且可以在培养中维持和扩增。它们对间充质干细胞标志物呈阳性,但对造血细胞标志物和移植物抗宿主病(GVHD)相关细胞标志物呈阴性。丹参处理导致华通氏胶细胞发生深刻的形态变化。诱导后的间充质干细胞形成具有多个神经突样延伸的圆形细胞体。最终它们形成了类似于神经元原代培养物的网络状突起。丹参和β-巯基乙醇还诱导间充质干细胞表达巢蛋白、β-微管蛋白III、神经丝(NF)和胶质纤维酸性蛋白(GFAP)。通过RT-PCR证实,间充质干细胞在丹参诱导前后均能表达多效生长因子。诱导后表达明显增强,并且巢蛋白基因也有表达。

结论

可以从人脐带华通氏胶中分离出间充质干细胞。使用丹参或β-巯基乙醇它们能够分化为神经样细胞。诱导后的间充质干细胞不仅发生形态变化,还表达神经元相关基因和神经元细胞标志物。它们可能代表中枢神经系统细胞移植的另一种干细胞来源。

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