In vitro isolation of stem cells derived from human dental pulp
In vitro isolation of stem cells derived from human dental pulp
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DOI：
10.1111/j.1399-0012.2009.01137.x
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发表时间：
2010-03-01
影响因子：
2.1
通讯作者：
Ali-Moghaddam, Kamran
中科院分区：
文献类型：
--
作者：
Agha-Hosseini, Farzaneh;Jahani, Mohammad-Ali;Ali-Moghaddam, Kamran
干细胞的特点是能够分化和自我更新。从人牙髓中提取的干细胞已被证明可以分化成成骨细胞，作为体外产生的自体骨的潜在来源。本研究的目的是从牙髓中分离间充质干细胞。本文对27例18-25岁的完整人恒磨牙进行牙髓轻轻拔除。用牛角钳在灭菌条件下分离完整牙髓。果肉在含有Dulbecco's modified Eagle's medium-low glucose (DMEM)-LG和Amphotericin 1%的培养基中培养。细胞随后传代扩增，两次传代后保存于液氮中作进一步检查。采用DMEM +胎牛血清（FBS） 10% l -谷氨酰胺0.1% +胰蛋白酶2.5% +乙二胺四乙酸（EDTA）传代。采用光镜和流式细胞术对细胞进行观察。分离的牙髓细胞表达间充质干细胞标记物。细胞中CD34、CD31和CD45呈阴性，而CD13、CD44、CD90、CD166和CD105呈阳性。这些结果表明，牙髓可以作为干细胞的来源，我们可以分离和培养。
Stem cells are characterized by the ability to differentiate and to self-renew. Stem cells derived from human dental pulp have been shown to differentiate into osteoblasts serving as a potential source of autologous bone produced in vitro. The purpose of the present study was to isolate mesenchymal stem cells from dental pulp. Dental pulp was gently extracted from 27 intact human permanent third molars of patients aged 18-25. Cow horn forceps were used to isolate intact dental pulp in sterilized condition. The pulps were cultured in a medium containing Dulbecco's modified Eagle's medium-low glucose (DMEM)-LG and Amphotericin 1%. The cells were subsequently expanded by passages, two passages were performed before they were stored in liquid nitrogen for further examination. DMEM + fetal bovine serum (FBS) 10% L-Glutamin 0.1% + Trypsin 2.5% + ethylene diamine tetraacetic acid (EDTA) were used for passage. Light microscope and flow cytometry were used to study the cells. The isolated dental pulp cells expressed mesenchymal stem cell markers. The cells were negative for CD34 and CD31 and CD45 but were positive for CD13, CD44, CD90, CD166, and CD105. These results indicate that dental pulp can be use as a source of stem cells that we can isolate and culture.