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纤连蛋白黏附、非纤连蛋白黏附及外植体来源的人牙髓干细胞群体的特性分析

Characterization of Fibronectin-Adherent, Non-Fibronectin-Adherent, and Explant-Derived Human Dental Pulp Stem Cell Populations.

作者信息

Kim Heoijin, Williams Shelley J, Colombo John S

机构信息

Department of Biomedical Sciences, School of Dental Medicine, University of Nevada, Las Vegas, NV 89106, USA.

出版信息

Dent J (Basel). 2025 Apr 2;13(4):159. doi: 10.3390/dj13040159.

Abstract

Dental pulp stem cells (DPSCs) are of significant interest due to their mesenchymal lineage and relative availability from extracted teeth. This study aims to examine the relationship between fibronectin-adherent, non-fibronectin-adherent, and explant-derived DPSC populations in terms of the population doubling rate in culture and the expression of mesenchymal cell surface markers and their capacity for osteodifferentiation. Human pulp tissue was removed from healthy extracted human teeth, enzymatically digested prior to seeding onto fibronectin-coated plates, and left to adhere for 20 min, yielding a fibronectin-adherent population. The remaining non-adherent cells were transferred and designated 'non-fibronectin-adherent.' Intact pulp was placed on uncoated plastic for 5 days, with the migrated cells designated 'explant-derived'. DPSCs from these populations were examined in terms of population doubling rates, the expression of CD90, CD44, CD105, and CD73, and the expression of , , and after 7 days in osteoinductive media. The fibronectin-adherent cells had the greatest population doubling over time. All populations demonstrated comparable percentages of cells positive for mesenchymal markers, though individual marker expression varied slightly. The explant-derived cells showed increased expression of after 7 days in osteoinductive media, while the treated single-cell-suspension-derived populations showed increased expression of mRNA. Fibronectin enrichment resulted in a population with the greatest rate of population doubling over extended culture compared to the other two populations. The proportion of cells positive for all four mesenchymal surface markers was the same between populations. The fibronectin-adherent and non-adherent cultures may have responded more rapidly to osteoinductive media than the explant-derived cells.

摘要

牙髓干细胞(DPSCs)因其间充质谱系以及从拔除牙齿中相对容易获取而备受关注。本研究旨在从培养中的群体倍增率、间充质细胞表面标志物的表达及其成骨分化能力方面,研究纤连蛋白黏附型、非纤连蛋白黏附型和外植体来源的DPSC群体之间的关系。从健康拔除的人牙中取出牙髓组织,在接种到纤连蛋白包被的培养板上之前进行酶消化,然后静置20分钟使其黏附,从而获得纤连蛋白黏附型群体。将剩余的未黏附细胞转移并命名为“非纤连蛋白黏附型”。将完整的牙髓置于未包被的塑料培养板上培养5天,将迁移出的细胞命名为“外植体来源型”。对这些群体来源的DPSCs进行群体倍增率、CD90、CD44、CD105和CD73表达的检测,并在成骨诱导培养基中培养7天后检测 、 和 的表达。随着时间推移,纤连蛋白黏附型细胞具有最大的群体倍增。所有群体间充质标志物阳性细胞的百分比相当,尽管个别标志物的表达略有差异。在成骨诱导培养基中培养7天后,外植体来源的细胞 表达增加,而经处理的单细胞悬液来源的群体 mRNA表达增加。与其他两个群体相比,纤连蛋白富集导致的群体在延长培养后具有最大的群体倍增率。各群体间四种间充质表面标志物阳性细胞的比例相同。纤连蛋白黏附型和非黏附型培养物对外植体来源的细胞相比,可能对成骨诱导培养基的反应更快。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1fc5/12026221/45c8364873e7/dentistry-13-00159-g0A1.jpg

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