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猫牙髓干细胞的分离与鉴定。

Isolation and characterization of feline dental pulp stem cells.

机构信息

Immunology and Immunotherapy Unit, Department of Patobiology, Veterinary Faculty, University of the Republic (UdelaR), Montevideo, Uruguay.

Odontostomatology Service, Veterinary Hospital Clinical Department, Veterinary Faculty, University of the Republic (UdelaR), Montevideo, Uruguay.

出版信息

J Feline Med Surg. 2023 Feb;25(2):1098612X221150625. doi: 10.1177/1098612X221150625.

Abstract

OBJECTIVES

The aim of this study was to isolate feline dental pulp stem cells (fDPSCs) and characterize their clonogenic and proliferative abilities, as well as their multipotency, immunophenotype and cytogenetic stability.

METHODS

Dental pulp was isolated by explant culture from two cats <1 year old at post mortem. Their clonogenicity was characterized using a colony-forming unit fibroblast assay, and their proliferative ability was quantified with a doubling time assay in passages 2, 4 and 6 (P, P and P, respectively). Multipotency was characterized with an in vitro trilineage differentiation assay in P, and cells were immunophenotyped in P by flow cytometry. Chromosomic stability was evaluated by cytogenetic analysis in P, P and P.

RESULTS

The fDPSCs displayed spindle and epithelial-like morphologies. Isolated cells showed a marked clonogenic capacity and doubling time was maintained from P to P. Trilineage differentiation was obtained in one sample, while the other showed osteogenic and chondrogenic differentiation. Immunophenotypic analysis showed fDPSCs were CD45, CD90 and CD44. Structural and numerical cytogenetic aberrations were observed in P-P.

CONCLUSIONS AND RELEVANCE

In this study, fDPSCs from two cats were isolated by explant culture and immunophenotyped. Cells displayed clonogenic and proliferative ability, and multipotency in vitro, and signs of chromosomic instability were observed. Although a larger study is needed to confirm these results, this is the first report of fDPSC isolation and in vitro characterization.

摘要

目的

本研究旨在分离猫牙髓干细胞(fDPSCs),并对其克隆形成和增殖能力,以及多能性、免疫表型和细胞遗传学稳定性进行鉴定。

方法

通过尸体剖检后<1 岁的两只猫的牙髓组织进行体外原代培养,以分离牙髓细胞。通过集落形成单位成纤维细胞检测鉴定其克隆形成能力,通过倍增时间试验分别在第 2 代(P2)、第 4 代(P4)和第 6 代(P6)量化其增殖能力。在 P 代,通过体外三系分化试验鉴定其多能性,并通过流式细胞术对 P 代细胞进行免疫表型鉴定。通过 P、P 和 P 代的细胞遗传学分析评估染色体稳定性。

结果

fDPSCs 呈纺锤形和上皮样形态。分离的细胞具有明显的克隆形成能力,倍增时间从 P2 保持到 P6。一个样本获得三系分化,而另一个样本则表现出成骨和成软骨分化。免疫表型分析显示 fDPSCs 表达 CD45、CD90 和 CD44。在 P-P 代观察到结构和数量染色体异常。

结论和相关性

在这项研究中,通过组织块培养和免疫表型鉴定从两只猫中分离出 fDPSCs。细胞表现出克隆形成和增殖能力,体外多能性,并且观察到染色体不稳定的迹象。尽管需要更大的研究来证实这些结果,但这是首次报道 fDPSC 的分离和体外鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3eeb/10812064/69ce2bccb5d6/10.1177_1098612X221150625-fig1.jpg

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