p75~(NTR)对人牙髓干细胞神经分化影响的初步研究
|
摘要
目的初步探索p75~(NTR)对人牙髓干细胞神经分化的影响。方法利用免疫磁珠分选的方法分选人牙髓干细胞,得到p75~(NTR)+细胞组,与牙髓干细胞未分选组的2组干细胞进行体外神经分化诱导。从细胞形态、RealTime PCR和Western blot蛋白印记检测Nestin、βⅢ-Tubulin、NSE的表达。结果体外神经诱导后DPSC胞体收缩,成球生长,形似神经元;Real-Time PCR和Western blot结果显示,两组细胞Nestin、βⅢ-Tubulin、NSE的表达均为阳性,其中牙髓干细胞未分选组神经分化效果最佳。结论 p75~(NTR)对人牙髓干细胞神经分化有抑制作用,机理还有待进一步研究。
Objective To investigate the effect of p75 neurotrophin receptor on the neural differention of dental pulp stem cells. Methods Dental pulp step cells were selected by magnetic activated cell sorting system( MACS) and identified by p75 neurotrophin receptor,and then they were incubated in the modified neural differentiation medium. The neuron-like cells were observed by microscope. Real-time RT-PCR and Western blot were used to detect the Nestin,βⅢ-Tubulin and NSE.Results All groups could different into neuron-like cells. The real-time RT-PCR and Western blot results showed that the expression of Nestin,βⅢ-Tubulin and NSE was positive after treatment. Conclusion P75 NTR could inhibite the neural differentiation of human dental pulp stem cells.
Objective To investigate the effect of p75 neurotrophin receptor on the neural differention of dental pulp stem cells. Methods Dental pulp step cells were selected by magnetic activated cell sorting system( MACS) and identified by p75 neurotrophin receptor,and then they were incubated in the modified neural differentiation medium. The neuron-like cells were observed by microscope. Real-time RT-PCR and Western blot were used to detect the Nestin,βⅢ-Tubulin and NSE.Results All groups could different into neuron-like cells. The real-time RT-PCR and Western blot results showed that the expression of Nestin,βⅢ-Tubulin and NSE was positive after treatment. Conclusion P75 NTR could inhibite the neural differentiation of human dental pulp stem cells.
引文
1 Gronthos S,Mankani M,Brahim J,et al.Postnatal human dental pulp stem cells(DPSCs)in vitro and in vivo.Proc Natl Acad Sci U S A,2000,97(25):13625-13630.
2 Janebodin K,Horst OV,Ieronimakis N,et al.Isolation and characterization of neural crest-derived stem cells from dental pulp of neonatal mice.PLo S One,2011,6(11):e27526.
3 王劲松,汪璇,王松灵.人乳牙牙髓干细胞体外表达和分泌胶质细胞源性神经营养因子的研究.北京口腔医学,2011,19(5):245-248.
4 Wang J,Wang X,Sun Z,et al.Stem cells from human-exfoliated deciduous teeth can differentiate into dopaminergic neuron-like cells.Stem Cells Dev,2010,19(9):1375-1383.
5 Matsushita K,Wang W,Itoh S,et al.Dental pulp can be a good candidate for nerve grafting in a xeno-graft model.J Neurosci Methods,2012,205(2):246-251.
6 Jarmalavicˇiūt e·A,Tunaitis V,Pivoraite·U,et al.Exosomes from dental pulp stem cells rescue human dopaminergic neurons from 6-hydroxy-dopamine-induced apoptosis.Cytotherapy,2015,17(7):932-939.
7 Chang CC,Chang KC,Tsai SJ,et al.Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media.J Formos Med Assoc,2014,113(12):956-965.
8 Alvarez-Viejo M,Menendez-Menendez Y,Otero-Hernandez J.CD271 as a marker to identify mesenchymal stem cells from diverse sources before culture.World J Stem Cells,2015,7(2):470-476.
9 Poloni A,Maurizi G,Rosini V,et al.Selection of CD271(+)cells and human AB serum allows a large expansion of mesenchymal stromal cells from human bone marrow.Cytotherapy,2009,11(2):153-162.
10 鞠迎新,温秀杰,杨琨,等.p75神经营养受体在胎鼠颌突外胚间充质干细胞体外矿化过程中表达变化的研究.中华口腔医学杂志,2016,51(7):426-431.
11 Yang K,Wang Y,Ju Y,et al.p75 neurotrophin receptor regulates differential mineralization of rat ectomesenchymal stem cells.Cell Prolif,2017,50(1).
12 Quirici N,Soligo D,Bossolasco P,et al.Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies.Exp Hematol,2002,30(7):783-791.
13 Battula VL,Treml S,Bareiss PM,et al.Isolation of functionally distinct mesenchymal stem cell subsets using antibodies against CD56,CD271,and mesenchymal stem cell antigen-1.Haematologica,2009,94(2):173-184.
14 Xing Y,Nie X,Chen G,et al.Comparison of P75 NTR-positive and-negative etcomesenchymal stem cell odontogenic differentiation through epithelial-mesenchymal interaction.Cell Prolif,2016,49(2):185-194.
15 Tomellini E,Lagadec C,Polakowska R,et al.Role of p75neurotrophin receptor in stem cell biology:more than just a marker.Cell Mol Life Sci,2014,71(13):2467-2481.
16 Mikami Y,Ishii Y,Watanabe N,et al.CD271/p75(NTR)inhibits the differentiation of mesenchymal stem cells into osteogenic,adipogenic,chondrogenic,and myogenic lineages.Stem Cells Dev,2011,20(5):901-913.
17 Waddington RJ,Youde SJ,Lee CP,et al.Isolation of distinct progenitor stem cell populations from dental pulp.Cells Tissues Organs,2009,189(1-4):268-274.
2 Janebodin K,Horst OV,Ieronimakis N,et al.Isolation and characterization of neural crest-derived stem cells from dental pulp of neonatal mice.PLo S One,2011,6(11):e27526.
3 王劲松,汪璇,王松灵.人乳牙牙髓干细胞体外表达和分泌胶质细胞源性神经营养因子的研究.北京口腔医学,2011,19(5):245-248.
4 Wang J,Wang X,Sun Z,et al.Stem cells from human-exfoliated deciduous teeth can differentiate into dopaminergic neuron-like cells.Stem Cells Dev,2010,19(9):1375-1383.
5 Matsushita K,Wang W,Itoh S,et al.Dental pulp can be a good candidate for nerve grafting in a xeno-graft model.J Neurosci Methods,2012,205(2):246-251.
6 Jarmalavicˇiūt e·A,Tunaitis V,Pivoraite·U,et al.Exosomes from dental pulp stem cells rescue human dopaminergic neurons from 6-hydroxy-dopamine-induced apoptosis.Cytotherapy,2015,17(7):932-939.
7 Chang CC,Chang KC,Tsai SJ,et al.Neurogenic differentiation of dental pulp stem cells to neuron-like cells in dopaminergic and motor neuronal inductive media.J Formos Med Assoc,2014,113(12):956-965.
8 Alvarez-Viejo M,Menendez-Menendez Y,Otero-Hernandez J.CD271 as a marker to identify mesenchymal stem cells from diverse sources before culture.World J Stem Cells,2015,7(2):470-476.
9 Poloni A,Maurizi G,Rosini V,et al.Selection of CD271(+)cells and human AB serum allows a large expansion of mesenchymal stromal cells from human bone marrow.Cytotherapy,2009,11(2):153-162.
10 鞠迎新,温秀杰,杨琨,等.p75神经营养受体在胎鼠颌突外胚间充质干细胞体外矿化过程中表达变化的研究.中华口腔医学杂志,2016,51(7):426-431.
11 Yang K,Wang Y,Ju Y,et al.p75 neurotrophin receptor regulates differential mineralization of rat ectomesenchymal stem cells.Cell Prolif,2017,50(1).
12 Quirici N,Soligo D,Bossolasco P,et al.Isolation of bone marrow mesenchymal stem cells by anti-nerve growth factor receptor antibodies.Exp Hematol,2002,30(7):783-791.
13 Battula VL,Treml S,Bareiss PM,et al.Isolation of functionally distinct mesenchymal stem cell subsets using antibodies against CD56,CD271,and mesenchymal stem cell antigen-1.Haematologica,2009,94(2):173-184.
14 Xing Y,Nie X,Chen G,et al.Comparison of P75 NTR-positive and-negative etcomesenchymal stem cell odontogenic differentiation through epithelial-mesenchymal interaction.Cell Prolif,2016,49(2):185-194.
15 Tomellini E,Lagadec C,Polakowska R,et al.Role of p75neurotrophin receptor in stem cell biology:more than just a marker.Cell Mol Life Sci,2014,71(13):2467-2481.
16 Mikami Y,Ishii Y,Watanabe N,et al.CD271/p75(NTR)inhibits the differentiation of mesenchymal stem cells into osteogenic,adipogenic,chondrogenic,and myogenic lineages.Stem Cells Dev,2011,20(5):901-913.
17 Waddington RJ,Youde SJ,Lee CP,et al.Isolation of distinct progenitor stem cell populations from dental pulp.Cells Tissues Organs,2009,189(1-4):268-274.