Oral Medicine Department/Dental Research Center, Faculty of Dentistry, Tehran University of Medical Sciences, Tehran, Iran.
Clin Transplant. 2010 Mar-Apr;24(2):E23-8. doi: 10.1111/j.1399-0012.2009.01137.x. Epub 2009 Dec 18.
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.
干细胞的特征是能够分化和自我更新。已经证明，从人牙髓中分离出的干细胞可以分化为成骨细胞，成为体外产生自体骨的潜在来源。本研究的目的是从牙髓中分离间充质干细胞。从 27 名年龄在 18-25 岁的患者的 27 颗完整的第三磨牙中轻轻提取牙髓。使用牛角钳在无菌条件下分离完整的牙髓。将牙髓在含有 Dulbecco 的改良 Eagle 培养基低糖（DMEM-LG）和两性霉素 1%的培养基中培养。细胞随后通过传代进行扩增，在传代两次后将其储存在液氮中以备进一步检查。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）用于传代。使用相差显微镜和流式细胞术研究细胞。分离的牙髓细胞表达间充质干细胞标志物。细胞对 CD34 和 CD31 和 CD45 呈阴性，但对 CD13、CD44、CD90、CD166 和 CD105 呈阳性。这些结果表明，牙髓可以作为我们可以分离和培养的干细胞的来源。
