不同保存介质对人少突胶质前体细胞生物学特性的影响
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摘要
背景:保存介质直接影响干细胞活性以及细胞功能,目前国内外缺乏对人少突胶质前体细胞保存介质方面的研究。目的:评价临床常用的保存介质对人少突胶质前体细胞活率的影响。方法:在4℃条件下,将处于对数生长期的人少突胶质前体细胞分别在少突细胞完全培养基、含有1%白蛋白的生理盐水、生理盐水中放置24 h,比较不同保存介质中的细胞活性以及继续培养后的细胞增殖能力。结果与结论:①细胞冷藏24h进行锥虫蓝染色,少突细胞完全培养基组、含有1%白蛋白的生理盐水组、生理盐水组细胞活率分别为83%,68%和51%,少突细胞完全培养基组细胞活率最高,单纯生理盐水组细胞活率最低;②细胞冷藏后继续培养1代,3种保存介质中少突胶质前体细胞的扩增能力有逐渐降低的趋势,少突细胞完全培养基组细胞增殖能力最高,其次是含有1%白蛋白的生理盐水组,单纯生理盐水中保存的细胞无法继续增殖。
BACKGROUND: Preservation media exert direct roles in stem cell activity and cell function, and there is still a lack of research on the preservation medium of human oligodendrocyte precursor cells worldwide.OBJECTIVE: To evaluate the effect of clinically used preservation media on the viability of human oligodendrocyte precursor cells.METHODS: Human oligodendrocyte precursor cells in logarithmic growth phase were cultured in human oligodendrocyte precursor cell complete medium, normal saline containing 1% albumin or normal saline for 24 hours at 4 °C. We then compared cell viability in different preservation media and cell proliferation ability after continuous culture.RESULTS AND CONCLUSION:(1) Cell viability was counted by trypan blue staining 24 hours after cell storage. The results showed that the cell viabilities in the oligodenocyte complete medium group, the normal saline group containing 1% albumin group, and the normal saline group were 83%, 68%, and 51%, respectively. The cell viability was highest in the oligodenocyte complete medium group and lowest in the normal saline group. Cryopreserved cells were further cultured, and the cell proliferation ability was declined gradually, which was highest in the oligodenocyte complete medium group, followed by the normal saline containing 1% albumin, whereas the oligodendrocytes preserved in the normal saline could not continue to proliferate.
BACKGROUND: Preservation media exert direct roles in stem cell activity and cell function, and there is still a lack of research on the preservation medium of human oligodendrocyte precursor cells worldwide.OBJECTIVE: To evaluate the effect of clinically used preservation media on the viability of human oligodendrocyte precursor cells.METHODS: Human oligodendrocyte precursor cells in logarithmic growth phase were cultured in human oligodendrocyte precursor cell complete medium, normal saline containing 1% albumin or normal saline for 24 hours at 4 °C. We then compared cell viability in different preservation media and cell proliferation ability after continuous culture.RESULTS AND CONCLUSION:(1) Cell viability was counted by trypan blue staining 24 hours after cell storage. The results showed that the cell viabilities in the oligodenocyte complete medium group, the normal saline group containing 1% albumin group, and the normal saline group were 83%, 68%, and 51%, respectively. The cell viability was highest in the oligodenocyte complete medium group and lowest in the normal saline group. Cryopreserved cells were further cultured, and the cell proliferation ability was declined gradually, which was highest in the oligodenocyte complete medium group, followed by the normal saline containing 1% albumin, whereas the oligodendrocytes preserved in the normal saline could not continue to proliferate.
引文
[1]Domingues HS,Cruz A,Chan JR,et al.Mechanical plasticity during oligodendrocyte differentiation and myelination.Glia.2018;66(1):5-14.
[2]Mauney SA,Pietersen CY,Sonntag KC,et al.Differentiation of oligodendrocyte precursors is impaired in the prefrontal cortex in schizophrenia.Schizophr Res.2015;169(1-3):374-380.
[3]Yang QK,Xiong JX,Yao ZX.Neuron-NG2 cell synapses:novel functions for regulating NG2 cell proliferation and differentiation.Biomed Res Int.2013;2013:402843.
[4]Lee Y,Morrison BM,Li Y,et al.Oligodendroglia metabolically support axons and contribute to neurodegeneration.Nature.2012;487(7408):443-448.
[5]Shimizu T,Osanai Y,Ikenaka K.Oligodendrocyte-Neuron Interactions:Impact on Myelination and Brain Function.Neurochem Res.2018;43(1):190-194.
[6]Miller RH.Oligodendrocyte origins.Trends Neurosci.1996;19(3):92-96.
[7]Lin SC,Bergles DE.Synaptic signaling between neurons and glia.Glia.2004;47(3):290-298.
[8]Nishiyama A.Glial progenitor cells in normal and pathological states.Keio J Med.1998;47(4):205-208.
[9]Scolding NJ,Pasquini M,Reingold SC,et al.Cell-based therapeutic strategies for multiple sclerosis.Brain.2017;140(11):2776-2796.
[10]Assinck P,Duncan GJ,Plemel JR,et al.Myelinogenic Plasticity of Oligodendrocyte Precursor Cells following Spinal Cord Contusion Injury.J Neurosci.2017;37(36):8635-8654.
[11]Higman MA,Port JD,Beauchamp NJ Jr,et al.Reversible leukoencephalopathy associated with re-infusion of DMSOpreserved stem cells.Bone Marrow Transplant.2000;26(7):797-800.
[12]Arutyunyan IV,Strokova SО,MakarovАV,et al.DMSO-Free Cryopreservation of Human Umbilical Cord Tissue.Bull Exp Biol Med.2018;166(1):155-162.
[13]Yu NH,Chun SY,Ha YS,et al.Optimal Stem Cell Transporting Conditions to Maintain Cell Viability and Characteristics.Tissue Eng Regen Med.2018;15(5):639-647.
[14]Correia C,Koshkin A,Carido M,et al.Effective Hypothermic Storage of Human Pluripotent Stem Cell-Derived Cardiomyocytes Compatible With Global Distribution of Cells for Clinical Applications and Toxicology Testing.Stem Cells Transl Med.2016;5(5):658-669.
[15]Lu?nik Z,Bertolin M,Breda C,et al.Preservation of Ocular Epithelial Limbal Stem Cells:The New Frontier in Regenerative Medicine.Adv Exp Med Biol.2016;951:179-189.
[16]Van Wezel-Meijler G,Steggerda SJ,Leijser LM.Cranial ultrasonography in neonates:role and limitations.Semin Perinatol.2010;34(1):28-38.
[17]Back SA.White matter injury in the preterm infant:pathology and mechanisms.Acta Neuropathol.2017;134(3):331-349.
[18]Ma J,Bo SH,Lu XT,et al.Protective effects of carnosine on white matter damage induced by chronic cerebral hypoperfusion.Neural Regen Res.2016;11(9):1438-1444.
[19]Back SA,Luo NL,Borenstein NS,et al.Late oligodendrocyte progenitors coincide with the developmental window of vulnerability for human perinatal white matter injury.JNeurosci.2001;21(4):1302-1312.
[20]Back SA,Riddle A,McClure MM.Maturation-dependent vulnerability of perinatal white matter in premature birth.Stroke.2007;38(2 Suppl):724-730.
[21]Jin K,Xie L,Mao X,et al.Effect of human neural precursor cell transplantation on endogenous neurogenesis after focal cerebral ischemia in the rat.Brain Res.2011;1374:56-62.
[22]Tabakow P,Jarmundowicz W,Czapiga B,et al.Transplantation of autologous olfactory ensheathing cells in complete human spinal cord injury.Cell Transplant.2013;22(9):1591-1612.
[23]Huang H,Chen L,Xi H,et al.Fetal olfactory ensheathing cells transplantation in amyotrophic lateral sclerosis patients:a controlled pilot study.Clin Transplant.2008;22(6):710-718.
[24]Zhao K,Li R,Bi S,et al.Combination of mild therapeutic hypothermia and adipose-derived stem cells for ischemic brain injury.Neural Regen Res.2018;13(10):1759-1770.
[25]Huang H,Chen L.Commentary:Neurorestoratology:AConcept and Emerging Discipline in the Treatment of Neurological Disorders.CNS Neurol Disord Drug Targets.2016;15(5):522-525.
[26]黄红云,毛更生,陈琳.神经修复临床细胞治疗现状与展望[J].中华细胞与干细胞杂志(电子版),2017,7(3):162-167.
[27]Song CG,Zhang YZ,Wu HN,et al.Stem cells:a promising candidate to treat neurological disorders.Neural Regen Res.2018;13(7):1294-1304.
[28]Faulkner J,Keirstead HS.Human embryonic stem cell-derived oligodendrocyte progenitors for the treatment of spinal cord injury.Transpl Immunol.2005;15(2):131-142.
[29]Rao RC,Boyd J,Padmanabhan R,et al.Efficient serum-free derivation of oligodendrocyte precursors from neural stem cell-enriched cultures.Stem Cells.2009;27(1):116-125.
[30]Neri M,Maderna C,Ferrari D,et al.Robust generation of oligodendrocyte progenitors from human neural stem cells and engraftment in experimental demyelination models in mice.PLoS One.2010;5(4):e10145.
[31]Wang C,Luan Z,Yang Y,et al.High purity of human oligodendrocyte progenitor cells obtained from neural stem cells:suitable for clinical application.J Neurosci Methods.2015;240:61-66.
[32]中国医药生物技术协会.干细胞制剂制备质量管理自律规范[J].中国医药生物技术,2016,11(6):481-490.
[33]陈海丹.干细胞临床研究政策回顾和展望[J].自然辩证法通讯,2018,40(3):81-86.
[34]Mohamadnejad M,Alimoghaddam K,Mohyeddin-Bonab M,et al.Phase 1 trial of autologous bone marrow mesenchymal stem cell transplantation in patients with decompensated liver cirrhosis.Arch Iran Med.2007;10(4):459-466.
[35]Mohyeddin-Bonab M,Mohamad-Hassani MR,Alimoghaddam K,et al.Autologous in vitro expanded mesenchymal stem cell therapy for human old myocardial infarction.Arch Iran Med.2007;10(4):467-473.
[36]Venkataramana NK,Kumar SK,Balaraju S,et al.Open-labeled study of unilateral autologous bone-marrowderived mesenchymal stem cell transplantation in Parkinson's disease.Transl Res.2010;155(2):62-70.
[37]Wang D,Zhang F,Shen W,et al.Mesenchymal stem cell injection ameliorates the inducibility of ventricular arrhythmias after myocardial infarction in rats.Int J Cardiol.2011;152(3):314-320.
[38]Zhang F,Ren H,Shao X,et al.Preservation media,durations and cell concentrations of short-term storage affect key features of human adipose-derived mesenchymal stem cells for therapeutic application.PeerJ.2017;5:e3301.
[39]Wu YD,Li M,Liao X,et al.Effects of storage culture media,temperature and duration on human adipose?derived stem cell viability for clinical use.Mol Med Rep.2019;19(3):2189-2201.
[40]Robinson NJ,Picken A,Coopman K.Low temperature cell pausing:an alternative short-term preservation method for use in cell therapies including stem cell applications.Biotechnol Lett.2014;36(2):201-209.
[41]Petrenko Y,Chudickova M,Vackova I,et al.Clinically Relevant Solution for the Hypothermic Storage and Transportation of Human Multipotent Mesenchymal Stromal Cells.Stem Cells Int.2019;2019:5909524.
[42]Utheim OA,Lyberg T,Eidet JR,et al.Effect of Transportation on Cultured Limbal Epithelial Sheets for Worldwide Treatment of Limbal Stem Cell Deficiency.Sci Rep.2018;8(1):10502.
[43]Campbell LH,Taylor MJ,Brockbank KG.Survey of Apoptosis After Hypothermic Storage of a Pancreaticβ-Cell Line.Biopreserv Biobank.2016;14(4):271-278.
[44]Pless-Petig G,Rauen U.Serum-Free Cryopreservation of Primary Rat Hepatocytes in a Modified Cold Storage Solution:Improvement of Cell Attachment and Function.Biopreserv Biobank.2018;16(4):285-295.
[2]Mauney SA,Pietersen CY,Sonntag KC,et al.Differentiation of oligodendrocyte precursors is impaired in the prefrontal cortex in schizophrenia.Schizophr Res.2015;169(1-3):374-380.
[3]Yang QK,Xiong JX,Yao ZX.Neuron-NG2 cell synapses:novel functions for regulating NG2 cell proliferation and differentiation.Biomed Res Int.2013;2013:402843.
[4]Lee Y,Morrison BM,Li Y,et al.Oligodendroglia metabolically support axons and contribute to neurodegeneration.Nature.2012;487(7408):443-448.
[5]Shimizu T,Osanai Y,Ikenaka K.Oligodendrocyte-Neuron Interactions:Impact on Myelination and Brain Function.Neurochem Res.2018;43(1):190-194.
[6]Miller RH.Oligodendrocyte origins.Trends Neurosci.1996;19(3):92-96.
[7]Lin SC,Bergles DE.Synaptic signaling between neurons and glia.Glia.2004;47(3):290-298.
[8]Nishiyama A.Glial progenitor cells in normal and pathological states.Keio J Med.1998;47(4):205-208.
[9]Scolding NJ,Pasquini M,Reingold SC,et al.Cell-based therapeutic strategies for multiple sclerosis.Brain.2017;140(11):2776-2796.
[10]Assinck P,Duncan GJ,Plemel JR,et al.Myelinogenic Plasticity of Oligodendrocyte Precursor Cells following Spinal Cord Contusion Injury.J Neurosci.2017;37(36):8635-8654.
[11]Higman MA,Port JD,Beauchamp NJ Jr,et al.Reversible leukoencephalopathy associated with re-infusion of DMSOpreserved stem cells.Bone Marrow Transplant.2000;26(7):797-800.
[12]Arutyunyan IV,Strokova SО,MakarovАV,et al.DMSO-Free Cryopreservation of Human Umbilical Cord Tissue.Bull Exp Biol Med.2018;166(1):155-162.
[13]Yu NH,Chun SY,Ha YS,et al.Optimal Stem Cell Transporting Conditions to Maintain Cell Viability and Characteristics.Tissue Eng Regen Med.2018;15(5):639-647.
[14]Correia C,Koshkin A,Carido M,et al.Effective Hypothermic Storage of Human Pluripotent Stem Cell-Derived Cardiomyocytes Compatible With Global Distribution of Cells for Clinical Applications and Toxicology Testing.Stem Cells Transl Med.2016;5(5):658-669.
[15]Lu?nik Z,Bertolin M,Breda C,et al.Preservation of Ocular Epithelial Limbal Stem Cells:The New Frontier in Regenerative Medicine.Adv Exp Med Biol.2016;951:179-189.
[16]Van Wezel-Meijler G,Steggerda SJ,Leijser LM.Cranial ultrasonography in neonates:role and limitations.Semin Perinatol.2010;34(1):28-38.
[17]Back SA.White matter injury in the preterm infant:pathology and mechanisms.Acta Neuropathol.2017;134(3):331-349.
[18]Ma J,Bo SH,Lu XT,et al.Protective effects of carnosine on white matter damage induced by chronic cerebral hypoperfusion.Neural Regen Res.2016;11(9):1438-1444.
[19]Back SA,Luo NL,Borenstein NS,et al.Late oligodendrocyte progenitors coincide with the developmental window of vulnerability for human perinatal white matter injury.JNeurosci.2001;21(4):1302-1312.
[20]Back SA,Riddle A,McClure MM.Maturation-dependent vulnerability of perinatal white matter in premature birth.Stroke.2007;38(2 Suppl):724-730.
[21]Jin K,Xie L,Mao X,et al.Effect of human neural precursor cell transplantation on endogenous neurogenesis after focal cerebral ischemia in the rat.Brain Res.2011;1374:56-62.
[22]Tabakow P,Jarmundowicz W,Czapiga B,et al.Transplantation of autologous olfactory ensheathing cells in complete human spinal cord injury.Cell Transplant.2013;22(9):1591-1612.
[23]Huang H,Chen L,Xi H,et al.Fetal olfactory ensheathing cells transplantation in amyotrophic lateral sclerosis patients:a controlled pilot study.Clin Transplant.2008;22(6):710-718.
[24]Zhao K,Li R,Bi S,et al.Combination of mild therapeutic hypothermia and adipose-derived stem cells for ischemic brain injury.Neural Regen Res.2018;13(10):1759-1770.
[25]Huang H,Chen L.Commentary:Neurorestoratology:AConcept and Emerging Discipline in the Treatment of Neurological Disorders.CNS Neurol Disord Drug Targets.2016;15(5):522-525.
[26]黄红云,毛更生,陈琳.神经修复临床细胞治疗现状与展望[J].中华细胞与干细胞杂志(电子版),2017,7(3):162-167.
[27]Song CG,Zhang YZ,Wu HN,et al.Stem cells:a promising candidate to treat neurological disorders.Neural Regen Res.2018;13(7):1294-1304.
[28]Faulkner J,Keirstead HS.Human embryonic stem cell-derived oligodendrocyte progenitors for the treatment of spinal cord injury.Transpl Immunol.2005;15(2):131-142.
[29]Rao RC,Boyd J,Padmanabhan R,et al.Efficient serum-free derivation of oligodendrocyte precursors from neural stem cell-enriched cultures.Stem Cells.2009;27(1):116-125.
[30]Neri M,Maderna C,Ferrari D,et al.Robust generation of oligodendrocyte progenitors from human neural stem cells and engraftment in experimental demyelination models in mice.PLoS One.2010;5(4):e10145.
[31]Wang C,Luan Z,Yang Y,et al.High purity of human oligodendrocyte progenitor cells obtained from neural stem cells:suitable for clinical application.J Neurosci Methods.2015;240:61-66.
[32]中国医药生物技术协会.干细胞制剂制备质量管理自律规范[J].中国医药生物技术,2016,11(6):481-490.
[33]陈海丹.干细胞临床研究政策回顾和展望[J].自然辩证法通讯,2018,40(3):81-86.
[34]Mohamadnejad M,Alimoghaddam K,Mohyeddin-Bonab M,et al.Phase 1 trial of autologous bone marrow mesenchymal stem cell transplantation in patients with decompensated liver cirrhosis.Arch Iran Med.2007;10(4):459-466.
[35]Mohyeddin-Bonab M,Mohamad-Hassani MR,Alimoghaddam K,et al.Autologous in vitro expanded mesenchymal stem cell therapy for human old myocardial infarction.Arch Iran Med.2007;10(4):467-473.
[36]Venkataramana NK,Kumar SK,Balaraju S,et al.Open-labeled study of unilateral autologous bone-marrowderived mesenchymal stem cell transplantation in Parkinson's disease.Transl Res.2010;155(2):62-70.
[37]Wang D,Zhang F,Shen W,et al.Mesenchymal stem cell injection ameliorates the inducibility of ventricular arrhythmias after myocardial infarction in rats.Int J Cardiol.2011;152(3):314-320.
[38]Zhang F,Ren H,Shao X,et al.Preservation media,durations and cell concentrations of short-term storage affect key features of human adipose-derived mesenchymal stem cells for therapeutic application.PeerJ.2017;5:e3301.
[39]Wu YD,Li M,Liao X,et al.Effects of storage culture media,temperature and duration on human adipose?derived stem cell viability for clinical use.Mol Med Rep.2019;19(3):2189-2201.
[40]Robinson NJ,Picken A,Coopman K.Low temperature cell pausing:an alternative short-term preservation method for use in cell therapies including stem cell applications.Biotechnol Lett.2014;36(2):201-209.
[41]Petrenko Y,Chudickova M,Vackova I,et al.Clinically Relevant Solution for the Hypothermic Storage and Transportation of Human Multipotent Mesenchymal Stromal Cells.Stem Cells Int.2019;2019:5909524.
[42]Utheim OA,Lyberg T,Eidet JR,et al.Effect of Transportation on Cultured Limbal Epithelial Sheets for Worldwide Treatment of Limbal Stem Cell Deficiency.Sci Rep.2018;8(1):10502.
[43]Campbell LH,Taylor MJ,Brockbank KG.Survey of Apoptosis After Hypothermic Storage of a Pancreaticβ-Cell Line.Biopreserv Biobank.2016;14(4):271-278.
[44]Pless-Petig G,Rauen U.Serum-Free Cryopreservation of Primary Rat Hepatocytes in a Modified Cold Storage Solution:Improvement of Cell Attachment and Function.Biopreserv Biobank.2018;16(4):285-295.