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牙髓干细胞与牙周干细胞的比较分析:形态、功能、成骨分化及蛋白质组学差异

Comparative Analysis of Dental Pulp and Periodontal Stem Cells: Differences in Morphology, Functionality, Osteogenic Differentiation and Proteome.

作者信息

Kotova Anastasia V, Lobov Arseniy A, Dombrovskaya Julia A, Sannikova Valentina Y, Ryumina Nadezhda A, Klausen Polina, Shavarda Alexey L, Malashicheva Anna B, Enukashvily Natella I

机构信息

Institute of Cytology of the Russian Academy of Sciences, 194064 St. Petersburg, Russia.

Cell Technologies Laboratory, General Dentistry Department, North-Western State Medical University, 191015 St. Petersburg, Russia.

出版信息

Biomedicines. 2021 Nov 3;9(11):1606. doi: 10.3390/biomedicines9111606.

Abstract

Dental stem cells are heterogeneous in their properties. Despite their common origin from neural crest stem cells, they have different functional capacities and biological functions due to niche influence. In this study, we assessed the differences between dental pulp stem cells (DPSC) and periodontal ligament stem cells (PDLSC) in their pluripotency and neuroepithelial markers transcription, morphological and functional features, osteoblast/odontoblast differentiation and proteomic profile during osteogenic differentiation. The data were collected in paired observations: two cell cultures, DPSC and PDLSC, were obtained from each donor. Both populations had the mesenchymal stem cells surface marker set exposed on their membranes but differed in (a marker of neuroectodermal origin) expression, morphology, and proliferation rate. mRNA was revealed in DPSC and PDLSC, while OCT4 protein was present in the nuclei of DPSC only. However, transcription of mRNA was 1000-10,000-fold lower in dental stem cells than in blastocysts. DPSC proliferated at a slower rate and have a shape closer to polygonal but they responded better to osteogenic stimuli as compared to PDLSC. mRNA was detected by qPCR in both types of dental stem cells but RUNX2 protein was detected by LC-MS/MS shotgun proteomics only in PDLSC suggesting the posttranscriptional regulation. and , marker genes of odontoblastic type of osteogenic differentiation, were transcribed in DPSC but not in PDLSC samples. Our results prove that DPSC and PDLSC are different in their biology and therapeutic potential: DPSC are a good candidate for osteogenic or odontogenic bone-replacement cell-seeded medicines, while fast proliferating PDLSC are a prospective candidate for other cell products.

摘要

牙干细胞在特性上具有异质性。尽管它们都共同起源于神经嵴干细胞,但由于微环境的影响,它们具有不同的功能能力和生物学功能。在本研究中,我们评估了牙髓干细胞(DPSC)和牙周膜干细胞(PDLSC)在多能性、神经上皮标志物转录、形态和功能特征、成骨细胞/成牙本质细胞分化以及成骨分化过程中的蛋白质组学特征方面的差异。数据收集于配对观察中:从每个供体获取两种细胞培养物,即DPSC和PDLSC。这两种细胞群体在细胞膜上都表达间充质干细胞表面标志物,但在(神经外胚层起源的标志物)表达、形态和增殖速率方面存在差异。在DPSC和PDLSC中均检测到mRNA,而OCT4蛋白仅存在于DPSC的细胞核中。然而,牙干细胞中mRNA的转录水平比囊胚中的低1000 - 10000倍。DPSC增殖速率较慢,形状更接近多边形,但与PDLSC相比,它们对成骨刺激的反应更好。通过qPCR在两种类型的牙干细胞中均检测到mRNA,但仅通过LC-MS/MS鸟枪法蛋白质组学在PDLSC中检测到RUNX2蛋白,这表明存在转录后调控。和成牙本质细胞型成骨分化的标志物基因在DPSC中转录,但在PDLSC样本中未转录。我们的结果证明,DPSC和PDLSC在生物学和治疗潜力方面存在差异:DPSC是成骨或成牙本质骨替代细胞接种药物的良好候选者,而快速增殖的PDLSC是其他细胞产品的潜在候选者。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1d74/8616025/51f0b9c7902c/biomedicines-09-01606-g001.jpg

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