Luke Alexander M, Patnaik Rajashree, Kuriadom Sam, Abu-Fanas Salem, Mathew Simy, Shetty Krishna P
College of Dentistry, Ajman University, Ajman, 346, United Arab Emirates.
Heliyon. 2020 Jan 6;6(1):e03054. doi: 10.1016/j.heliyon.2019.e03054. eCollection 2020 Jan.
Human dental pulp stem cells (hDPSCs) are promising source of cells for numerous and varied regenerative medicine applications as those possess high proliferation potential with multilineage differentiation capacity compare to other sources of adult stem cells; therefore, hDPSCs could be the good source for autologous transplantation in tissue engineering and regenerative medicine. In this study stem cells were isolated from dental pulp and were characterised by flowcytometry and immunocytochemistry. The controlled cells as well as, 7-day cultured cells were positive for transcription factors, OCT 4 and SOX 2 thatconfirmed isolated cellsasmesenchymal stem cells (MSCs). These cells showed positive expression for CD 19, CD 73, CD 90, CD 105 and are negative for CD 34, CD 45. Viability of hDPSCS were studied by trypan blue (TB) staining and fluorescent microscopic study. After 7 days of passaging by using several growth factors, cells express neural cell markers oligodendrocyte and glial fibrillary acidic protein. Specifically, osteocytes were grown from dental pulp MSCSs with the help of growth factors, dexamethasone, ascorbic acid-2- phosphate and β-glycerophosphate whereas, adipocytes were grown with indomethacin, 3-isobutyl-1-methylxanthine and insulin. Osteocytes and adipocytes were characterized by von Kossa and Oil red O staining, respectively. Chromosomal analysis of dental pulp-MSCs was done for qualitative assessment of MSCs. Karyotyping indicated diploid chromosome number in dental pulp derived MSCs. grown osteocytes could be used for bone fracture reunion cases, and adipocytes could be used for further research purposes.
人牙髓干细胞（hDPSCs）是用于众多不同再生医学应用的有前景的细胞来源，因为与其他成体干细胞来源相比，它们具有高增殖潜力和多谱系分化能力；因此，hDPSCs可能是组织工程和再生医学中自体移植的良好来源。在本研究中，从牙髓中分离出干细胞，并通过流式细胞术和免疫细胞化学进行表征。对照细胞以及培养7天的细胞对转录因子OCT 4和SOX 2呈阳性，这证实分离出的细胞为间充质干细胞（MSCs）。这些细胞对CD 19、CD 73、CD 90、CD 105呈阳性表达，对CD 34、CD 45呈阴性表达。通过台盼蓝（TB）染色和荧光显微镜研究来研究hDPSCS的活力。在使用几种生长因子传代7天后，细胞表达神经细胞标志物少突胶质细胞和胶质纤维酸性蛋白。具体而言，在生长因子地塞米松、抗坏血酸-2-磷酸酯和β-甘油磷酸酯的帮助下，从牙髓MSCSs中培养出骨细胞，而在吲哚美辛、3-异丁基-1-甲基黄嘌呤和胰岛素的作用下培养出脂肪细胞。分别通过冯·科萨染色和油红O染色对骨细胞和脂肪细胞进行表征。对牙髓-MSCs进行染色体分析以对MSCs进行定性评估。核型分析表明牙髓来源的MSCs中染色体数目为二倍体。培养的骨细胞可用于骨折愈合病例，脂肪细胞可用于进一步的研究目的。
