深低温冻存对牙周膜干细胞膜片增殖和分化能力的影响

›› 2015, Vol. 6 ›› Issue (2): 87-89.

深低温冻存对牙周膜干细胞膜片增殖和分化能力的影响

魏福兰¹,马晓妮²,张凡³

1. 山东大学口腔医院正畸科
2. 山东大学口腔医院种植科
3. 山东大学口腔医院正畸科

收稿日期:2015-03-27 修回日期:2015-06-03 出版日期:2015-06-25 发布日期:2015-07-13
通讯作者: 魏福兰 E-mail:fulanwei777@126.com

Effect of cryopreservation on proliferation and differentiation of periodontal ligament stem cell sheet

Received:2015-03-27 Revised:2015-06-03 Online:2015-06-25 Published:2015-07-13

摘要/Abstract

摘要： 目的：研究深低温冻存对牙周膜干细胞膜片增殖和分化能力的影响。方法：将牙周膜干细胞膜片用90%胎牛血清+10% DMSO冻存液冻存3月后复苏，采用甲基噻唑基四唑（MTT）法、von Kossa染色和油红O染色法检测冻存对增殖和分化的影响。结果：冻存牙周膜干细胞膜片与新鲜牙周膜干细胞膜片的增殖率和成骨、成脂分化能力没有明显差异。结论：牙周膜干细胞膜片经深低温保存后可继续保持冻存前原有的增殖能力和多向分化能力。

Abstract: Objective：To investigate the effect of cryopreservation on proliferation and differentiation of periodontal ligament stem cells (PDLSCs) sheet. Methods：We compared cryopreserved PDLSCs sheet with freshly PDLSCs sheet in terms of cell proliferation rate and multilineage differentiation potential by MTT, von Kossa staining and Oil Red staining. Results：Cryopreserved PDLSCs sheet showed a similar cell proliferation rate and multilineage differentiation potential as fresh PDLSCs sheet. Conclusions：This study demonstrates that cryopreservation does not affect the proliferation and differentiation of PDLSCs sheet, supporting the feasibility of PDLSCs sheet cryopreservation in nitrogen.

魏福兰马晓妮张凡. 深低温冻存对牙周膜干细胞膜片增殖和分化能力的影响[J]. , 2015, 6(2): 87-89.

参考文献

[1]Ding G, Liu Y, Wang W, et al.Allogeneic periodontal ligament stem cell therapy for periodontitis in swine[J].Stem Cells,2010,28(10):1829-1838.
[2]Wei F, Qu C, Song T, et al.Vitamin C treatment promotes mesenchymal stem cell sheet formation and t9 regeneration by elevating telomerase activity[J].J Cell Physiol,2012,227(9):3216-3224.
[3]Khammanit R, Chantakru S, Kitiyanant Y, et al.Effect of serum starvation and chemical inhibitors on cell cycle synchronization of canine dermal fibroblasts[J].Theriogenology,2008,70(1):27-34.
[4]de Barros FR, Goissis MD, Caetano HV, et al.Serum starvation and full confluency for cell cycle synchronization of domestic cat (felis catus) foetal fibroblasts[J].Reprod Domest Anim,2010,45(1):38-41.
[5]Iwata T, Yamato M, Tsuchioka H, et al.Periodontal regeneration with multi-layered periodontal ligament-derived cell sheets in a canine model[J].Biomaterials,2009,30(14):2716-2723.
[6]Flores MG, Hasegawa M, Yamato M, et al.Cementum-periodontal ligament complex regeneration using the cell sheet technique[J].J Periodontal Res,2008,43(3):364-371.
[7]Seo BM, Miura M, Gronthos S, et al.Investigation of multipotent postnatal stem cells from human periodontal ligament[J].Lancet,2004,364(9429):149-155.
[8]Yang Z, Jin F, Zhang X, et al.T4 engineering of cementum/periodontal-ligament complex using a novel three-dimensional pellet cultivation system for human periodontal ligament stem cells[J].T4 Eng Part C Methods,2009,15(4):571-581.
[9]Shin JS, Hong SW, Lee SL, et al.Serum starvation induces G1 arrest through suppression of Skp2-CDK2 and CDK4 in SK-OV-3 cells[J].Int J Oncol,2008,32(2):435-439.
[10]Alfieri R, Barberis M, Chiaradonna F, et al.Towards a systems biology approach to mammalian cell cycle: modeling the entrance into S phase of quiescent fibroblasts after serum stimulation[J].BMC Bioinformatics,2009,10 (Suppl 12):S16-.
[11]Miranda Mdos S, Bressan FF, Zecchin KG, et al.Serum-starved apoptotic fibroblasts reduce blastocyst production but enable development to term after SCNT in cattle[J].Cloning Stem Cells,2009,11(4):565-573.
[12]Fellah BH, Josselin N, Chappard D, et al.Inflammatory reaction in rats muscle after implantation of biphasic calcium phosphate micro particles[J].J Mater Sci Mater Med,2007,18(2):287-294.
[13]Goissis MD, Caetano HV, Marques MG, et al.Effects of serum deprivation and cycloheximide on cell cycle of low and high passage porcine fetal fibroblasts[J].Reprod Domest Anim,2007,42 (6):660-663.
[14]Akahane M, Shigematsu H, Tadokoro M, et al.Scaffold-free cell sheet injection results in bone formation[J].J T5 Eng Regen Med,2010,4(5):404-411.
[15]Nakamura A, Akahane M, Shigematsu H, et al.Cell sheet transplantation of cultured mesenchymal stem cells enhances bone formation in a rat nonunion model[J].Bone,2010,46(2):418-424.
[16]Bianchi V, Coticchio G, Fava L, et a1.Meiotic spindle imaging in human oocytes frozen with a slow freezing procedure involving high sucrose concentration[J].Hum Reprod,2005,20(4):1078-1083.
[17]Gorlin J.Stem cell cryopreservation[J].J Infus Chemother,1996,6(1):23-27.
[18]Bakken AM.Cryopreserving human peripheral blood progenitor cells[J].Curr Stem Cell Res Ther,2006,1(1):47-54.