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从患有 Legg-Calvé-Perthes 病的犬受影响的股骨头中生成和鉴定间充质干细胞。

Generation and characterization of mesenchymal stem cells from the affected femoral heads of dogs with Legg Calvé Perthes disease.

机构信息

Laboratory of Veterinary Surgery, Department of Veterinary Medicine, College of Bioresource and Sciences, Nihon University, Fujisawa, Japan.

出版信息

Open Vet J. 2024 May;14(5):1172-1181. doi: 10.5455/OVJ.2024.v14.i5.12. Epub 2024 May 31.

Abstract

BACKGROUND

Canine Legg Calvé Perthes disease (LCPD) occurs during the growth period, and the cause of ischemic necrosis of the femoral head during growth remains unclear. If LCPD-affected femoral head-derived mesenchymal stem cells (LCPD-MSCs) can be generated, they can be used as a new tool for the pathophysiological analysis of canine LCPD.

AIM

To generate affected femoral head-derived mesenchymal stem cells (MSCs) from dogs with LCPD and investigate the mRNA expression levels of angiogenesis-related factors and osteogenic differentiation potency of LCPD-MSCs.

METHODS

This study was performed using affected femoral heads from dogs diagnosed with LCPD and underwent femoral head and neck ostectomy. The necrotic tissue was harvested from the LCPD-affected femoral head and cultured statically (LCPD group, = 6). Canine bone marrow-derived MSCs (BM-MSCs) were used as controls (control group, = 6). First, the morphology of the cultured cells was observed, and the expression of CD29, CD34, CD44, CD45, CD90, and major histocompatibility complex class II was analyzed using flow cytometry. Additionally, the trilineage differentiation potency of the LCPD-affected head-derived adherent cells was examined. Furthermore, the expression levels of , , , and mRNAs and the bone differentiation potency of LCPD-affected head-derived adherent cells were investigated.

RESULTS

LCPD-affected femoral head-derived adherent cells showed a fibroblast-like morphology, and the expression of cell surface antigens was similar to that of BM-MSCs. In addition, LCPD-affected femoral head-derived adherent cells showed the same trilineage differentiation potency as BM-MSCs and were consistent with MSC characteristics. Furthermore, the mRNA expression levels of angiogenesis-related factors could be objectively measured in LCPD-MSCs and those MSCs had bone differentiation potency.

CONCLUSION

In the present study, canine LCPD-MSCs were successfully generated, suggesting their usefulness as a tool for pathological analysis of LCPD in dogs.

摘要

背景

犬 Legg-Calvé-Perthes 病(LCPD)发生于生长期,而生长期股骨头缺血性坏死的原因尚不清楚。如果能生成 LCPD 病变股骨头来源的间充质干细胞(LCPD-MSCs),则可将其作为犬 LCPD 病理生理学分析的新工具。

目的

从 LCPD 犬的病变股骨头中生成病变股骨头来源的间充质干细胞(MSCs),并研究 LCPD-MSCs 中与血管生成相关的因子的 mRNA 表达水平和成骨分化潜能。

方法

本研究使用 LCPD 犬接受股骨头颈切除术的病变股骨头。从 LCPD 病变股骨头中采集坏死组织,进行静态培养(LCPD 组,n=6)。以犬骨髓来源的 MSCs(BM-MSCs)作为对照(对照组,n=6)。首先观察培养细胞的形态,并用流式细胞术分析 CD29、CD34、CD44、CD45、CD90 和主要组织相容性复合体 II 的表达。此外,检测 LCPD 病变头来源的贴壁细胞的三系分化潜能。进一步研究 LCPD 病变头来源的贴壁细胞中 、 、 和 mRNA 的表达水平及骨分化潜能。

结果

LCPD 病变股骨头来源的贴壁细胞呈成纤维细胞样形态,细胞表面抗原的表达与 BM-MSCs 相似。此外,LCPD 病变股骨头来源的贴壁细胞具有与 BM-MSCs 相同的三系分化潜能,符合 MSC 特征。进一步研究发现,LCPD-MSCs 中可客观测量到与血管生成相关因子的 mRNA 表达水平,且这些 MSC 具有骨分化潜能。

结论

本研究成功生成了犬 LCPD-MSCs,提示其可用作犬 LCPD 病理分析的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/65a6/11199743/633d88aaa5c4/OpenVetJ-14-1172-g001.jpg

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