Human umbilical cord mesenchymal stem cells co-cultured with dental pulp cells in vitro and its effect on cell biological behaviors

doi:10.19439/j.sjos.2018.04.006

Abstract

Abstract: PURPOSE: To investigated the effect of human umbilical cord mesenchymal stem cells (hUCMSCs) and human dental pulp cells (hDPCs) on cell biological behaviors by co-culture system in vitro. METHODS: hUCMSCs and hDPCs were obtained by primary culture. A culture system of hUCMSCs and hDPCs induced by BMP2 was established in vitro. hUCMSCs and hDPCs were co-cultured at the ratio of 1:1, 1:5 and 5:1. The optimum ratio of each group was selected to further experiment. The formation of calcium nodule was stained by alizarin red staining at 21 day. The expression of DSPP,ALP,DMP1,OCN,VEGF,HGF and Nanog gene was detected by real-time quantitative PCR at 7 day and 14 day. 1:1 group and hUCMSCs, hDPCs group were selected for alizarin red staining at 21 day according to PCR results. Statistical analysis was performed using SPSS 21.0 software package. RESULTS: Calcified nodules formation in 1:1 group was significantly higher than in hUCMSCs group (P<0.05), close to that in hDPCs. qPCR showed that the mRNA expression of DSPP, ALP, DMP1, OCN, VEGF and HGF in 1:1 group was significantly higher than that in hUCMSCs (P<0.05); mRNA expression of Nanog in 1:1 group was significantly lower than in hUCMSCs group (P<0.05). The results of alizarin red staining showed that the OD value of 1:1 group was significantly higher than that of hUCMSCs group (P<0.05). CONCLUSIONS: The cells can be induced to differentiate into odontoblastoid-like cells and the mRNA expression of angiogenic factors was stimulated by hUCMSCs co-culure wih hDPCs.

Key words: Human umbilical cord mesenchymal stem cells, Human dental pulp cells, Co-culture

CLC Number:

R781.3

LIN Tian, ZHAO Wen-qing, LU Yan-ling, LIU Yu-ying, BAO Li-rong, WU Yu. Human umbilical cord mesenchymal stem cells co-cultured with dental pulp cells in vitro and its effect on cell biological behaviors[J]. Shanghai Journal of Stomatology, 2018, 27(4): 365-369.

References

[1] Wu LF, Wang NN, Liu YS, et al.Differentiation of Wharton's jelly primitive stromal cells into insulin-producing cells in comparison with bone marrow mesenchymal stem cells[J]. Tissue Eng Part A, 2009, 15(10): 2865-2873.
[2] Seshareddy K, Troyer D, Weiss ML.Method to isolate mesenchymal-like cells from Wharton's Jelly of umbilical cord[J]. Methods Cell Biol, 2008, 86: 101-119.
[3] Zhang YN, Lie PC, Wei X.Differentiation of mesenchymal stromal cells derived from umbilical cord Wharton's jelly into hepatocyte-like cells[J]. Cytotherapy, 2009, 11(5): 548-558.
[4] De Bruyn C, Najar M, Raicevic, et al. A rapid, simple, and reproducible method for the isolation of mesenchymal stromal cells from Wharton's jelly without enzymatic treatment[J]. Stem Cells Dev, 2011, 20(3): 547-557.
[5] Hartmann I, Hollweck T, Haffner S, et al.Umbilical cord tissue-derived mesenchymal stem cells grow best under GMP-compliant culture conditions and maintain their phenotypic and functional properties[J]. J Immunol Methods, 2010, 363(1): 80-89.
[6] Li TX, Yuan J, Chen Y, et al.Differentiation of mesenchymal stem cells from human umbilical cord tissue into odontoblast-like cells using the conditioned medium of tooth germ cells in vitro[J]. Biomed Res Int, 2013, 2013: 218543.
[7] 赵文青, 林田, 陆彦玲, 等. 骨诱导形成蛋白2诱导牙髓细胞矿化最佳条件的探索[J]. 广西医科大学学报, 2015, 32(6): 879-883.
[8] 林田, 陆彦玲, 包丽荣, 等. 人脐带间充质干细胞培养体系的优化[J]. 广西医科大学学报, 2017, 34(4): 500-504.
[9] Hsieh JY, Fu SJ, Chang SJ, et al.Functional module analysis reveals differential osteogenic and stemness potentials in human mesenchymal stem cells from bone marrow and Wharton's jelly of umbilical cord[J]. Stem Cells Dev, 2010, 19(12): 1895-1910.
[10] Gnecchi M, Zhang Z, Ni A, et al.Paracrine mechanisms in adult stem cell signaling and therapy[J]. Circ Res, 2008, 103(11): 1204-1219.
[11] Mao C, Hou X, Wang B, et al.Intramuscular injection of human umbilical cord-derived mesenchymal stem cells improves cardiac function in dilated cardiomyopathy rats[J]. Stem Cell Res Ther, 2017, 8(1): 18.
[12] Joggerst SJ, Hatzopoulos AK.Stem cell therapy for cardiac repair: benefits and barriers[J]. Expert Rev Mol Med, 2009, 11: e20.
[13] Nagaya N, Kangawa K, Itoh T, et al.Transplantation of mesenchymal stem cells improves cardiac function in a rat model of dilated cardiomyopathy[J]. Circulation, 2005, 112(8): 1128-1135.
[14] Zhang J, Liu X, Yu W, et al.Effects of human Vascular endothelial growth factor on reparative dentin formation[J]. Mol Med Rep, 2016, 13(1): 705-712.
[15] Tabata MJ, Kim K, Liu JG, et al.Hepatocyte growth factor is involved in the morphogenesis of tooth germ in murine molars[J]. Development, 1996, 122(4): 1243-1251.
[16] Yuan C, Wang P, Zhu l, et al. Coculture of stem cells from apical papilla and human umbilical vein endothelial cell under hypoxia increases the formation of three-dimensional vessel-like structures in vitro[J]. Tissue Eng Part A, 2015, 21(5-6): 1163-1172.
[17] Wang J, Rao S, Chu J, et al.A protein interaction network for pluripotency of embryonic stem cells[J]. Nature, 2006, 444(7117): 364-368.
[18] Liang J, Wan M, Zhang Y, et al.Nanog and Oct4 associate with unique transcriptional repression compiexes in embryonic stem cells[J]. Nat Cell Biol, 2008, 10(6): 731-739.