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猫子宫内膜间充质干细胞的分离与鉴定。

Isolation and characterization of feline endometrial mesenchymal stem cells.

机构信息

Department of Veterinary Internal Medicine, College of Veterinary Medicine, Chungnam National University, Daejeon 34134, Korea.

CM Animal Hospital, Jincheon 27802, Korea.

出版信息

J Vet Sci. 2024 Mar;25(2):e31. doi: 10.4142/jvs.23267.

Abstract

BACKGROUND

Recently, there has been a growing interest in stem cells for human medicine. Limited feline endometrial mesenchymal stem cell (fEM-MSC) research in veterinary medicine necessitates reporting for future feline disease research and therapy.

OBJECTIVES

This study aimed to isolate fEM-MSCs from feline endometrial tissues and evaluate their morphology, proliferative ability, differentiation ability, and immunophenotype.

METHODS

Feline endometrial tissues were obtained from the ovariohysterectomies of healthy cats and isolated using an enzymatic method. The morphology and proliferative ability of the isolated cells were assessed using a doubling time (DT) assay from passages 3 to 6 (P3 - P6). We measured pluripotency gene expressions of cells in P2 using quantitative real-time polymerase chain reaction (qRT-PCR). To investigate MSC characteristics, a trilineage differentiation assay was conducted in P4, and cells in P4 were immunophenotyped using flow cytometry.

RESULTS

fEM-MSCs showed a typical spindle-shaped morphology under a microscope, and the DT was maintained from P3 to P6. fEM-MSCs could differentiate into adipocytes, osteoblasts, and chondrocytes, and expressed three pluripotency markers (OCT4, SOX2, and NANOG) by qRT-PCR. Immunophenotypic analysis showed that the fEM-MSCs were CD14 , CD34 , CD45 , CD9, and CD44.

CONCLUSIONS

In this study, the feline endometrium was a novel source of MSCs, and to the best of our knowledge, this is the first report on the isolation method and characteristics of fEM-MSCs.

摘要

背景

最近,人们对人类医学中的干细胞产生了越来越大的兴趣。兽医领域对有限的猫子宫内膜间充质干细胞(fEM-MSC)的研究需要进行报告,以便为未来的猫疾病研究和治疗提供参考。

目的

本研究旨在从猫子宫内膜组织中分离 fEM-MSCs,并评估其形态、增殖能力、分化能力和免疫表型。

方法

从健康猫的卵巢子宫切除术获得猫子宫内膜组织,并使用酶法进行分离。使用倍增时间(DT)测定法(P3-P6)评估分离细胞的形态和增殖能力。我们使用定量实时聚合酶链反应(qRT-PCR)在 P2 测量细胞的多能性基因表达。为了研究 MSC 特征,在 P4 进行三系分化测定,并用流式细胞术对 P4 中的细胞进行免疫表型分析。

结果

fEM-MSCs 在显微镜下呈典型的纺锤形形态,从 P3 到 P6 保持 DT。fEM-MSCs 可分化为脂肪细胞、成骨细胞和软骨细胞,并通过 qRT-PCR 表达三个多能性标记物(OCT4、SOX2 和 NANOG)。免疫表型分析显示 fEM-MSCs 表达 CD14、CD34、CD45、CD9 和 CD44。

结论

在本研究中,猫子宫内膜是 MSC 的新来源,据我们所知,这是首次报道 fEM-MSCs 的分离方法和特征。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a3ed/10990916/0dc756414d32/jvs-25-e31-g001.jpg

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