Mehrabani Davood, Mahdiyar Parisa, Torabi Kianoosh, Robati Reza, Zare Shahrokh, Dianatpour Mehdi, Tamadon Amin
Assistant Professor of Stem Cell and Transgenic Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran; Assistant Professor of Department of Developmental Biology, Science and Research Branch, Islamic Azad University, Fars, Iran; Assistant Professor of Department of Regenerative Medicine, University of Manitoba, Winnipeg, Manitoba, Canada.
Assistant Professor of Stem Cell and Transgenic Technology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
J Clin Exp Dent. 2017 Feb 1;9(2):e172-e177. doi: 10.4317/jced.52824. eCollection 2017 Feb.
Dental pulp stem cells (DPSCs) play an important role in tissue regeneration. This study compares the growth kinetics and characterization of third molar and first premolar human DPSCs.
Dental pulp tissues were isolated from human first premolar and third molar teeth and were digested by treating them with collagenase type I. Single-cell suspensions from each dental pulp were seeded in T25 culture flasks and the media were replaced every 3 days until 70% confluence. The cells were enumerated to determine the population doubling time (PDT). Cells were characterized using flow cytometry, RT-PCR and osteogenic medium for differentiation of DPSCs. Karyotyping assay was also performed till passage 7th.
The DPSCs had spindle-shaped morphology. There was an increase in PDT in third molar DPSCs when compared to first premolar teeth. Positive expression of CD44, CD73, and CD90 and negative expression of CD34 and CD45 were illustrated. A normal karyotype was visible for all seven passages. The Alizarin red staining was positive for osteogenic induction of DPSCs.
When DPSCs are needed, third molar teeth can be a good and convenient candidate for cell transplantation, yielding high number of cells with mesenchymal characteristics. They can be a source for further investigations and work on tissue engineering protocols. Stem cells, dental pulp, growth kinetics, characterization.
牙髓干细胞(DPSCs)在组织再生中发挥着重要作用。本研究比较了人类第三磨牙和第一前磨牙牙髓干细胞的生长动力学和特性。
从人类第一前磨牙和第三磨牙中分离牙髓组织,并用I型胶原酶进行消化处理。将每个牙髓的单细胞悬液接种于T25培养瓶中,每3天更换一次培养基,直至细胞汇合度达到70%。对细胞进行计数以确定群体倍增时间(PDT)。使用流式细胞术、逆转录-聚合酶链反应(RT-PCR)以及用于牙髓干细胞分化的成骨培养基对细胞进行特性分析。直至第7代还进行了核型分析检测。
牙髓干细胞呈纺锤形形态。与第一前磨牙牙髓干细胞相比,第三磨牙牙髓干细胞的群体倍增时间有所增加。结果显示CD44、CD73和CD90呈阳性表达,CD34和CD45呈阴性表达。所有七代细胞均可见正常核型。茜素红染色显示牙髓干细胞的成骨诱导呈阳性。
当需要牙髓干细胞时,第三磨牙可成为细胞移植的良好且便利的供体,能产生大量具有间充质特性的细胞。它们可作为进一步研究以及组织工程方案研究的细胞来源。干细胞、牙髓、生长动力学、特性分析。