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人牙髓干细胞神经源性分化潜能的免疫表型及分子分析

Immunophenotypic and Molecular Analysis of Human Dental Pulp Stem Cells Potential for Neurogenic Differentiation.

作者信息

Fatima Nikhat, Khan Aleem A, Vishwakarma Sandeep K

机构信息

Department of Periodontics, Pacific University, Udaipur, Rajasthan, India.

Central Laboratory for Stem Cell Research and Translational Medicine, CLRD, Deccan College of Medical Sciences, Kanchanbagh, Hyderabad, Telangana, India.

出版信息

Contemp Clin Dent. 2017 Jan-Mar;8(1):81-89. doi: 10.4103/ccd.ccd_998_16.

Abstract

BACKGROUND

Growing evidence shows that dental pulp (DP) tissues could be a potential source of adult stem cells for the treatment of devastating neurological diseases and several other conditions.

AIMS

Exploration of the expression profile of several key molecular markers to evaluate the molecular dynamics in undifferentiated and differentiated DP-derived stem cells (DPSCs) .

SETTINGS AND DESIGN

The characteristics and multilineage differentiation ability of DPSCs were determined by cellular and molecular kinetics. DPSCs were further induced to form adherent (ADH) and non-ADH (NADH) neurospheres under serum-free condition which was further induced into neurogenic lineage cells and characterized for their molecular and cellular diversity at each stage.

STATISTICAL ANALYSIS USED

Statistical analysis used one-way analysis of variance, Student's -test, Livak method for relative quantification, and R programming.

RESULTS

Immunophenotypic analysis of DPSCs revealed >80% cells positive for mesenchymal markers CD90 and CD105, >70% positive for transferring receptor (CD71), and >30% for chemotactic factor (CXCR3). These cells showed mesodermal differentiation also and confirmed by specific staining and molecular analysis. Activation of neuronal lineage markers and neurogenic growth factors was observed during lineage differentiation of cells derived from NADH and ADH spheroids. Greater than 80% of cells were found to express β-tubulin III in both differentiation conditions.

CONCLUSIONS

The present study reported a cascade of immunophenotypic and molecular markers to characterize neurogenic differentiation of DPSCs under serum-free condition. These findings trigger the future analyses for clinical applicability of DP-derived cells in regenerative applications.

摘要

背景

越来越多的证据表明,牙髓(DP)组织可能是用于治疗严重神经疾病和其他几种病症的成体干细胞的潜在来源。

目的

探索几种关键分子标志物的表达谱,以评估未分化和分化的牙髓来源干细胞(DPSCs)中的分子动态。

设置与设计

通过细胞和分子动力学确定DPSCs的特征和多向分化能力。在无血清条件下,将DPSCs进一步诱导形成贴壁(ADH)和非贴壁(NADH)神经球,再将其进一步诱导分化为神经谱系细胞,并对每个阶段的分子和细胞多样性进行表征。

所用统计分析方法

采用单因素方差分析、学生t检验、Livak相对定量法和R编程进行统计分析。

结果

DPSCs的免疫表型分析显示,>80%的细胞间充质标志物CD90和CD105呈阳性,>70%的细胞转铁蛋白受体(CD71)呈阳性,>30%的细胞趋化因子(CXCR3)呈阳性。这些细胞也表现出中胚层分化,通过特异性染色和分子分析得到证实。在源自NADH和ADH球体的细胞谱系分化过程中,观察到神经谱系标志物和神经源性生长因子的激活。在两种分化条件下,均发现>80%的细胞表达β-微管蛋白III。

结论

本研究报道了一系列免疫表型和分子标志物,用于表征无血清条件下DPSCs的神经源性分化。这些发现引发了对牙髓来源细胞在再生应用中临床适用性的进一步分析。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/06cb/5426172/1b920974986b/CCD-8-81-g001.jpg

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