人骨髓基质细胞的体内骨形成与体外迁移能力之间的关联。

Association between in vivo bone formation and ex vivo migratory capacity of human bone marrow stromal cells.

作者信息

Andersen Rikke K, Zaher Walid, Larsen Kenneth H, Ditzel Nicholas, Drews Katharina, Wruck Wasco, Adjaye James, Abdallah Basem M, Kassem Moustapha

机构信息

Department of Endocrinology and Metabolism, University Hospital of Odense, Odense, Denmark.

Stem Cell Unit, Department of Anatomy, College of Medicine, King Saud University, Riyadh, Saudi Arabia.

出版信息

Stem Cell Res Ther. 2015 Oct 8;6:196. doi: 10.1186/s13287-015-0188-9.

DOI:10.1186/s13287-015-0188-9

PMID:26450135

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4599318/

Abstract

INTRODUCTION

There is a clinical need for developing systemic transplantation protocols for use of human skeletal stem cells (also known bone marrow stromal stem cells) (hBMSC) in tissue regeneration. In systemic transplantation studies, only a limited number of hBMSC home to injured tissues suggesting that only a subpopulation of hBMSC possesses "homing" capacity. Thus, we tested the hypothesis that a subpopulation of hBMSC defined by ability to form heterotopic bone in vivo, is capable of homing to injured bone.

METHODS

We tested ex vivo and in vivo homing capacity of a number of clonal cell populations derived from telomerized hBMSC (hBMSC-TERT) with variable ability to form heterotopic bone when implanted subcutaneously in immune deficient mice. In vitro transwell migration assay was used and the in vivo homing ability of transplanted hBMSC to bone fractures in mice was visualized by bioluminescence imaging (BLI). In order to identify the molecular phenotype associated with enhanced migration, we carried out comparative DNA microarray analysis of gene expression of hBMSC-derived high bone forming (HBF) clones versus low bone forming (LBF) clones.

RESULTS

HBF clones were exhibited higher ex vivo transwell migration and following intravenous injection, better in vivo homing ability to bone fracture when compared to LBF clones. Comparative microarray analysis of HBF versus LBF clones identified enrichment of gene categories of chemo-attraction, adhesion and migration associated genes. Among these, platelet-derived growth factor receptor (PDGFR) α and β were highly expressed in HBF clones. Follow up studies showed that the chemoattractant effects of PDGF in vitro was more enhanced in HBF compared to LBF clones and this effect was reduced in presence of a PDGFRβ-specific inhibitor: SU-16 f. Also, PDGF exerted greater chemoattractant effect on PDGFRβ(+) cells sorted from LBF clones compared to PDGFRβ(-) cells.

CONCLUSION

Our data demonstrate phenotypic and molecular association between in vivo bone forming ability and migratory capacity of hBMSC. PDGFRβ can be used as a potential marker for the prospective selection of hBMSC populations with high migration and bone formation capacities suitable for clinical trials for enhancing bone regeneration.

摘要

引言

临床上需要开发系统性移植方案，以便在组织再生中使用人类骨骼干细胞（也称为骨髓基质干细胞）（hBMSC）。在系统性移植研究中，只有有限数量的hBMSC归巢至受损组织，这表明只有hBMSC的一个亚群具有“归巢”能力。因此，我们检验了这样一个假设，即体内形成异位骨能力所定义的hBMSC亚群能够归巢至受损骨骼。

方法

我们测试了源自端粒酶逆转录酶转导的hBMSC（hBMSC-TERT）的多个克隆细胞群体的体外和体内归巢能力，这些克隆细胞群体在免疫缺陷小鼠皮下植入时形成异位骨的能力各不相同。采用体外transwell迁移试验，并通过生物发光成像（BLI）观察移植的hBMSC在小鼠体内向骨折部位的归巢能力。为了鉴定与增强迁移相关的分子表型，我们对hBMSC来源的高骨形成（HBF）克隆与低骨形成（LBF）克隆的基因表达进行了比较DNA微阵列分析。

结果

与LBF克隆相比，HBF克隆表现出更高的体外transwell迁移能力，静脉注射后，其在体内向骨折部位的归巢能力更强。对HBF克隆与LBF克隆进行比较微阵列分析，确定了趋化吸引、黏附及迁移相关基因类别的富集。其中，血小板衍生生长因子受体（PDGFR）α和β在HBF克隆中高表达。后续研究表明，与LBF克隆相比，PDGF在体外对HBF克隆的趋化吸引作用更强，且在存在PDGFRβ特异性抑制剂SU-16 f的情况下，这种作用减弱。此外，与PDGFRβ（-）细胞相比，PDGF对从LBF克隆中分选的PDGFRβ（+）细胞的趋化吸引作用更强。