CN113564122A - 人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用 - Google Patents
人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用 Download PDFInfo
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- CN113564122A CN113564122A CN202110894940.XA CN202110894940A CN113564122A CN 113564122 A CN113564122 A CN 113564122A CN 202110894940 A CN202110894940 A CN 202110894940A CN 113564122 A CN113564122 A CN 113564122A
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Abstract
本发明涉及生物技术领域,具体涉及人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用。所述方法包括使用以下至少一种培养基培养干细胞:神经诱导完全培养基、N2培养基、B27培养基、OPC成熟培养基;更优选的,使用含有葛根素的OPC成熟培养基诱导少突胶质细胞相较于不使用葛根素可以提高30%少突胶质细胞的数量。
Description
技术领域
本发明涉及生物技术领域,具体涉及人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用。
背景技术
少突胶质前体细胞(Oligodendrocyte progenitor cells,OPC)是存在于脊椎动物中的中枢神经系统细胞,它形成髓鞘包绕神经纤维,为神经信号的传递和神经纤维的营养及保护起关键作用。少突胶质细胞产生富含脂质的层状髓鞘,分化成熟形成髓鞘,其覆盖神经元轴突并产生限定的电绝缘区段以使动作电位传导速度最大化。髓磷脂对于轴突完整性和存活也是重要的,并且已显示即使影响少突胶质细胞代谢的小变化也可导致神经退化。髓鞘化过程在人类中特别重要,因为人脑具有较高含量的有髓鞘神经元(白质),并且髓鞘化在出生之后持续并贯穿一生。这表明少突胶质细胞不只提供惰性绝缘,而且髓鞘化是动态过程,从而影响认知功能乃至行为。多发性硬化症(MS)、肾上腺脑白质营养不良、白质消融性疾病、佩利措伊斯-梅茨巴赫病(Pelizaeus-Merzbacher disease)和脑白质营养不良是脱髓鞘或髓鞘形成障碍病症的实例。此外,少突胶质细胞的关键作用正显现在许多其它神经病症和神经退化性病状,包括肌萎缩性侧索硬化、亨廷顿氏病(Huntington’sdisease)、阿尔茨海默氏病(Alzheimer's disease)和精神分裂症中。
神经干细胞生物学和临床应用的进展为难以治愈的神经系统疾病的治疗提供了可能性。同时,提高被植入的OPC的存活率以及改善它们的微环境对于利用神经干细胞治疗神经系统疾病也具有至关重要的作用。来源于人的OPC不仅有助于更好的了解少突胶质细胞的功能及轴突-神经元的相互作用,更为髓鞘的修复研究及药物治疗提供了必不可少的工具。
人诱导性多能干细胞(iPSC)可被体外诱导成神经干细胞(NSC),进而产生不同谱系的神经元、胶质细胞,如少突胶质前体细胞(OPCs),或星形胶质细胞。iPSC也可被直接诱导成OPCs,但目前技术所需的时间长,且效率低。
发明内容
本发明提供了一种由小分子和其他因子介导的,经诱导性多能干细胞向少突胶质细胞分化的改良方法,本发明基于相同的iPSC经NSC诱导OPCs的改良方法,并且,实验中证实了葛根素增加经诱导性多能干细胞向少突胶质细胞分化的诱导效率和数量,在诱导后期可增加30%少突胶质细胞的数量。
方法
第一方面,本发明提供了一种诱导少突胶质细胞的方法,所述方法包括使用以下培养基至少一种培养干细胞:
1)神经诱导完全培养基;
2)N2培养基;
3)B27培养基;
4)OPC成熟培养基。
优选地,所述方法是依次使用神经诱导完全培养基、N2培养基、B27培养基、OPC成熟培养基进行干细胞培养。
优选地,使用所述神经诱导完全培养基的时间是4-10天;优选地,7天;更优选地,第0-7天。
优选地,使用所述N2培养基的时间是1-6天;优选地,3天;更优选地,第8-11天。
优选地,使用所述B27培养基的时间是4-10天;优选地,7天;更优选地,第12-19天。
优选地,使用所述OPC成熟培养基的时间是至少7天;优选地,至少10天;更优选地,第20-30天。
优选地,所述的培养是对来源于人、猩猩、猴、马、牛、羊、猪、驴、骆驼、狗、兔、猫、大鼠、小鼠、鱼、鸟或昆虫的任意细胞进行的细胞培养。
优选地,所述干细胞包括ESC(胚胎干细胞)、诱导性多能干细胞(iPSC)、拟胚体、细胞造血干细胞、神经干细胞、间充质干细胞、皮肤干细胞、脂肪干细胞、脐血干细胞中的一种或多种。
更优选地,所述干细胞是iPSC。
优选地,所述iPSC细胞可以是商品化的细胞系,也可以是由供体细胞诱导而来,所述供体细胞包括绒毛细胞、皮肤(成纤维细胞和角质形成细胞)、羊水、胚外组织(胎盘和脐带)、脐带血、骨膜、牙组织、脂肪组织、神经干细胞、肝细胞、间质干细胞、外周血细胞、乳腺上皮细胞、脂肪干细胞,脐带基质和胎盘中的一种或多种。
在一种实施方式中,所述神经诱导完全培养基中含有第一小分子化合物组合。
在一种实施方式中,所述神经诱导完全培养基是由第一基础培养基、非必须氨基酸、谷氨酰胺、还原剂和第一小分子化合物组合组成。
优选地,本发明所述的第一基础培养基、第二基础培养基、第三基础培养基、第四基础培养基各自独立地包括TeSR-E8、mTESR1、E8、Essential 8TMMedium、达尔伯克改良伊格尔培养基(DMEM,Dulbecco’s Modified Eagle’s Medium)、最小必需培养基(MEM,MinimalEssential Medium)、伊格尔基本培养基(BME,Basal Medium Eagle)、F-10、F-12、α-最小必需培养基(α-MEM,α-Minimal Essential Medium)、G-最小必需培养基(G-MEM、Glasgow’sMinimal Essential Medium)、IMPM(IMDM,Iscove’s Modified Dulbecco’s Medium)、AmnioMax、新型二代羊水培养基(Amino MaxⅡcomplete Medium,Gibco,Newyork,USA)、Chang’s培养基、Mesem Cult-XF培养基(STEMCELL Technologies,Vancouver,Canada)、RPMI1640、Ham’s F12、DMEM/F12、Ham's F-12K Medium、Hepato ZYME-SFM、William’s EMedium、Waymouth’s Medium或Hepatocyte Culture Medium。
优选地,本发明所述第一基础培养基、第二基础培养基、第三基础培养基、第四基础培养基是DMEM/F12。
优选地,本发明所述还原剂包括但不限于β-巯基乙醇(2-Mercaptoethanol)、二硫苏糖醇、二硫赤藓糖醇、还原型谷胱甘肽、半胱氨酸、硫代胺甲酸盐、二硫亚磺酸钠、抗坏血酸盐、二氯化锡或硼氢化钠。
优选地,本发明所述谷氨酰胺选用GlutaMAX-I,所述“GlutaMAX”是一种细胞培养添加剂,可直接替代细胞培养基中的L-谷氨酰胺。
优选地,本发明所述非必须氨基酸包括丙氨酸、精氨酸、天门冬氨酸、胱氨酸、脯氨酸、酪氨酸。
优选地,所述神经诱导完全培养基是由98%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、2-Mercaptoethanol和第一小分子化合物组合组成。
优选地,所述神经诱导完全培养基是由98%DMEM/F-12培养基(LifeTechnologies,cat.no.11039021)、1%非必须氨基酸(Life Technologies,cat.no.11140-050)、1%GlutaMAX-I(Life Technologies,cat.no.35050061)、2-Mercaptoethanol(Gibco,31350010)和第一小分子化合物组合组成。“括号”内代表所述产品的厂家及货号。
优选地,所述第一小分子化合物组合包括胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂中的至少一种。
优选地,所述TGF-β信号通路抑制剂包括SB431542、Dorsomorphin、A83-01、LDN193189、RepSox、SB 525334、DMH-1、SB-505124、BMP signaling agonist sb4、GW788388、ITD-1、LSKL,SD-208、LDN-212854、K02288、LDN-214117、R-268712、SM16、A 77-01、ALK2-IN-2、PD-161570、pm26、TGF-β1peptide TFA、Isosaponarin、BIO-013077-01。
优选地,所述TGF-β信号通路抑制剂选用SB431542;优选地,其货号为medchemexpress,HY-10431。
优选地,所述SB431542的工作浓度为10μM。
优选地,所述BMP信号通路抑制剂包括Dorsomorphin、A83-01、LDN193189、RepSox、SB 525334、DMH-1、SB-505124、BMP signaling agonist sb4、GW788388、ITD-1、LSKL、SD-208、LDN-212854、K02288、LDN-214117、R-268712、SM16、A 77-01、ALK2-IN-2、PD-161570、pm26、TGF-β1peptide TFA、Isosaponarin、BIO-013077-01。
优选地,所述BMP信号通路抑制剂是LDN193189;优选地,其货号为medchemexpress,HY-12071。
优选地,所述LDN193189的工作浓度为0.25μM。
优选地,所述RAR核受体的激动剂包括维生素A酸、Rapamycin、3-Methyladenine、Acetylcysteine、5-Fluorouracil,Hydrocortisone、Docetaxel、Rosiglitazone、Estradiol、Melatonin、GW9662、Nicotinamide、Cytarabine、Isoprenalinehydrochloride、Prostaglandin E2、Acetaminophen、(-)-Epigallocatechin Gallate、β-Nicotinamide mononucleotide、Calcitriol、DHEA、Liothyronine、NAD+、Luteolin、Thymidine、Docosahexaenoic Acid、Kaempferol、Palmitic acid、Cyclopamine、Genistein、L-Glutathione reduced。
优选地,所述RAR核受体的激动剂是维生素A酸;优选地,其货号为medchemexpress,HY-14649。
优选地,所述维生素A酸的工作浓度为100μM。
在一种实施方式中,所述N2培养基中包含第二小分子化合物组合。
在一种实施方式中,所述N2培养基是由第二基础培养基、非必须氨基酸、谷氨酰胺、还原剂、N2 supplement和第二小分子化合物组合组成。
本发明所述的“N2 supplement”中含有人全铁转铁蛋白(HumanTransferrinHolo)、重组胰岛素全链(InsulinRecombinantFullChain)、孕酮(Progesterone)、腐胺(Putrescine)、亚硒酸盐(Selenite),其可以是商品化的产品或自行配制获得。
优选地,所述N2培养基由97%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、2-Mercaptoethanol、1%N2 supplement和第二小分子化合物组合组成。
优选地,所述N2培养基由97%DMEM/F-12培养基(Life Technologies,cat.no.11039021)、1%非必须氨基酸(Life Technologies,cat.no.11140-050)、1%GlutaMAX-I(Life Technologies,cat.no.35050061)、2-Mercaptoethanol(Gibco,31350010)、1%N2 supplement(ThermoFisher,cat.no.17502001)和第二小分子化合物组合组成。“括号”内代表所述产品的厂家及货号。
优选地,所述第二小分子化合物组合包括RAR核受体的激动剂和/或Hedgehog信号通路激活剂。
优选地,所述RAR核受体的激动剂与前述一致。
优选地,所述Hedgehog信号通路激活剂包括SAG、Cyclopamine、Purmorphamine、20(S)-Hydroxycholesterol、Halcinonide、Jervine、ALLO-2、IHR-Cy3。
优选地,所述Hedgehog信号通路激活剂是SAG;优选地,其货号为medchemexpress,HY-12848。
在一种实施方式中,所述B27培养基中包含第三小分子化合物组合。
在一种实施方式中,所述B27培养基是由第三基础培养基、非必须氨基酸、谷氨酰胺、还原剂、N2 supplement、B27 supplement和第三小分子化合物组合组成。
本发明所述“B27 supplement”中含有生物素(Biotin)、DL-α-生育酚乙酸酯(DLAlpha Tocopherol Acetate)、DL-α-生育酚(DLAlpha-Tocopherol)、BSA(fattyacidfreeFractionV)、过氧化氢酶(Catalase)、人重组胰岛素(Human RecombinantInsulin)、人转铁蛋白(Human Transferrin)、超氧化物歧化酶(Superoxide Dismutase)、皮质酮(Corticosterone)、D-半乳糖(D-Galactose)、乙醇胺盐酸(Ethanolamine HCl)、还原型谷胱甘肽(Glutathione reduced)、左旋肉碱盐酸(L-Carnitine HCl)、亚油酸(Linoleic Acid)、亚麻酸(Linolenic Acid)、孕酮(Progesterone)、腐胺(Putrescine2HCl)、亚硒酸钠(Sodium Selenite)、三碘甲状腺原氨酸(T3 triodo-I-thyronine),其可以是商品化的产品或自行配制获得。
优选地,所述B27培养基由95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、2-Mercaptoethanol、1%N2 supplement、2%B27 supplement和第三小分子化合物组合组成。
优选地,所述B27培养基由95%DMEM/F-12培养基(Life Technologies,cat.no.11039021)、1%非必须氨基酸(Life Technologies,cat.no.11140-050)、1%GlutaMAX-I(Life Technologies,cat.no.35050061)、2-Mercaptoethanol(Gibco,31350010)、1%N2 supplement(ThermoFisher,cat.no.17502001)、2%B27supplement(ThermoFisher,cat.no.12587010)和第三小分子化合物组合组成。“括号”内代表所述产品的厂家及货号。
优选地,所述第三小分子化合物组合包括胰岛素、RAR核受体的激动剂和Hedgehog信号通路激活剂中的至少一种。
优选地,所述RAR核受体的激动剂和Hedgehog信号通路激活剂与前述一致。
在一种实施方式中,所述OPC成熟培养基中包含第四小分子化合物组合。
在一种实施方式中,所述OPC成熟培养基还包括第四基础培养基、非必须氨基酸、谷氨酰胺、还原剂、N2 supplement、B27 supplement。
优选地,所述OPC成熟培养基还包括95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、2-Mercaptoethanol、1%N2 supplement、2%B27supplement。
优选地,所述OPC成熟培养基还包括95%DMEM/F-12培养基(Life Technologies,cat.no.11039021)、1%非必须氨基酸(Life Technologies,cat.no.11140-050)、1%GlutaMAX-I(Life Technologies,cat.no.35050061)、2-Mercaptoethanol(Gibco,31350010)、1%N2 supplement(ThermoFisher,cat.no.17502001)、2%B27 supplement(ThermoFisher,cat.no.12587010)。“括号”内代表所述产品的厂家及货号。
优选地,所述第四小分子化合物组合包括胰岛素、PDGF-AA、IGF-1、HGF、NT3、T3、Biotin、cAMP中的至少一种。
优选地,所述第四小分子化合物组合包括胰岛素、PDGF-AA(PeproTech,cat.no.AF-100-13A)、IGF-1(PeproTech,cat.no.AF-100-11)、HGF(PeproTech,cat.no.100-39H)、NT3(PeproTech,cat.no.450-03)、T3(Sigma-Aldrich,cat.no.T2877)、Biotin(Sigma-Aldrich,cat.no.B4639)、cAMP(Sigma-Aldrich,cat.no.D0260)和5-HT2C受体拮抗剂。“括号”内代表所述产品的厂家及货号。
优选地,所述第四小分子化合物组合中还含有5-HT2C受体拮抗剂;
优选地,所述5-HT2C受体拮抗剂包括Puerarin(葛根素)、Harmine、SCH-23390hydrochloride、Olanzapine、GTS-21dihydrochloride、Thioridazinehydrochloride、Risperidone、SB-269970hydrochloride、Dihydroergotamine mesylate、Amitriptyline hydrochloride、Aripiprazole、Ketanserin、WAY-100635Maleate、Buspirone hydrochloride、Cisapride、Methiothepin mesylate、Trazodonehydrochloride、Pindolol、Quetiapine、Lumateperone tosylate、Perphenazine、8-OH-DPAT、Levomepromazine、SB-224289hydrochloride、Volinanserin、Ziprasidone、Paliperidone、Sertindole、Alprenolol hydrochloride、Asenapine maleate。
优选地,所述5-HT2C受体拮抗剂是Puerarin;优选地,其货号为medchemexpress,HY-N0145。
另一方面本发明提供了一种诱导神经干细胞的方法,所述方法包括前述神经诱导完全培养基和/或前述N2培养基进行细胞培养。
所述方法包括前述神经诱导完全培养基和前述N2培养基进行细胞培养。
小分子化合物组合
另一方面本发明还提供了一种包含5-HT2C受体拮抗剂的小分子化合物组合。
优选地,所述小分子化合物组合是前述第四小分子化合物组合。
试剂盒
另一方面本发明还提供了诱导少突胶质细胞的试剂盒,所述试剂盒中包括前述胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂、Hedgehog信号通路激活剂、5-HT2C受体拮抗剂中的一种或多种。
优选地,所述试剂盒中包括配置以下至少一种培养基的试剂:
1)神经诱导完全培养基;
2)N2培养基;
3)B27培养基;
4)OPC成熟培养基。
应用
另一方面,本发明提供了以下任意一种培养基及其在诱导少突胶质细胞中的应用:
1)神经诱导完全培养基;
2)N2培养基;
3)B27培养基;
4)OPC成熟培养基。
另一方面,本发明提供了包含5-HT2C受体拮抗剂的小分子化合物组合在诱导少突胶质细胞中的应用。
另一方面,本发明提供了Puerarin在诱导少突胶质细胞中的应用。
另一方面,本发明提供了前述试剂盒在诱导少突胶质细胞中的应用。
另一方面,本发明提供了胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂、Hedgehog信号通路激活剂、5-HT2C受体拮抗剂任意一种在诱导少突胶质细胞中的应用。
另一方面,本发明提供了胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂、Hedgehog信号通路激活剂在诱导神经干细胞中的应用。
细胞及其应用
另一方面,本发明提供了一种细胞群,其特征在于,所述细胞群选自以下一种:
1)由前述诱导神经干细胞的方法制备得到的细胞群,所述细胞群中包括表达NKX2.2+、Nestin+、PAX6+中至少一种的细胞。
优选地,所述细胞群中存在表达Olig2+的细胞。
2)由前述诱导少突胶质细胞的方法制备得到的细胞群,所述细胞群中少突胶质细胞占全部细胞比例的至少20%;优选地,至少21%、25%、30%、35%、36%。
优选地,所述少突胶质细胞是表达Olig2、Nkx2.2、O4和MBP中至少一种的细胞。
优选地,所述少突胶质细胞是Olig2和Nkx2.2共表达的细胞;或者,所述少突胶质细胞是O4和MBP共表达的细胞。
另一方面,本发明提供了经过前述诱导少突胶质细胞的方法制得的细胞在治疗中枢神经系统疾病中的用途。
优选地,所述细胞是少突胶质细胞。
优选地,所述中枢神经系统疾病包括但不限于神经退行性疾病、脱髓鞘疾病、癫痫、脑外伤、休克、痴呆、青光眼、神经系统损伤后再生以及精神类疾病。
优选地,所述神经退行性疾病包括阿尔兹海默症、小脑萎缩症、原发性侧索硬化、脊髓性肌萎缩症、帕金森病、亨廷顿氏病、克雅二氏病、牛海绵状脑病、共济失调毛细血管扩张症、肌肉萎缩性侧索硬化症、精神类疾病。
本发明所述“脱髓鞘疾病”是指髓鞘形成后发生的髓鞘损坏,脱髓鞘疾病是以神经髓鞘脱失为主,神经元胞体及轴突相对受累较轻为特征的一组疾病。
优选地,所述脱髓鞘疾病包括遗传性和获得性两大类。
优选地,所述脱髓鞘疾病包括脑卒中、多发性硬化、视神经脊髓炎、急性播散性脑脊髓炎、弥漫性硬化、同心圆硬化、脑白质营养不良、脑桥中央髓鞘溶解症、急性炎症性脱髓鞘性多发性神经病、慢性炎症性脱髓鞘性多发性神经病、缺血-缺氧性病引起的白质脑病、营养缺乏性疾病引起的亚急性联合变性、病毒感染引起的亚急性硬化性全脑炎或进行性多灶性白质脑病、糖尿病性神经病、系统性红斑狼疮的神经病变、肾上腺脑白质营养不良、白质消融性疾病、佩利措伊斯-梅茨巴赫病(Pelizaeus-Merzbacher disease)。
优选地,所述精神性疾病包括精神分裂症,抑郁症,偏执狂,焦虑症,强迫症,恐惧症,
另一方面,本发明提供了治疗前述神经疾病的方法,所述方法包括通过使用前述方法诱导少突胶质细胞。
附图说明
图1是第16天免疫荧光检测图;A:olig2+,Nkx2.2+,B:olig2+,Nestin+,C:olig2+,PAX6+。
图2是第33天免疫荧光检测图。
图3是OPC成熟培养基中添加或不添加葛根素对比得到的少突胶质细胞百分比统计结果图。
图4是OPC成熟培养基中添加或不添加葛根素对比的免疫荧光检测图。
具体实施方式
下面结合实施例对本发明做进一步的说明,以下所述,仅是对本发明的较佳实施例而已,并非对本发明做其他形式的限制,任何熟悉本专业的技术人员可能利用上述揭示的技术内容加以变更为同等变化的等效实施例。凡是未脱离本发明方案内容,依据本发明的技术实质对以下实施例所做的任何简单修改或等同变化,均落在本发明的保护范围内。
本发明所使用的试剂
*二抗:与一抗对应的Alexa Fluor荧光-标记抗体
通用方法:免疫荧光染色鉴定
工作液的配置:
1.配置10ml封闭血清稀释液(5%BSA+0.5%Triton X-100+DPBS液,以配置10ml为例。即在9.4ml DPBS中加入500μl正常5%BSA和100μl 30%Triton X-100)。
2.配置一抗工作液:封闭血清稀释液加入合适一抗滴度(见一抗使用说明书中具体滴度值)。
3.配置二抗工作液:封闭血清稀释液加入合适二抗滴度(见二抗使用说明书中具体滴度值)。
4.配置90%甘油:用DPBS稀释。
免疫荧光染色具体步骤如下:
DPBS清洗三遍,3min/次,4%PFA室温固定40min。DPBS清洗三遍,3min/次。0.5%TritonX-100,穿孔15min。5%BSA+0.15%TritonX-100,室温封闭1h。配PBST:DPBS+1%BSA+0.15%TritonX-100。上一抗,4度过夜。回收一抗溶液,PBST清洗三遍,每次10min。上二抗,1:500,4度过夜,避光。PBST清洗三遍,每次10min。5μg/ml DAPI for 2-3min,避光。用DPBS清洗一遍,加入90%的甘油。
实施例1、诱导iPSC细胞分化,并验证诱导效果
配制以下培养基备用:
神经诱导完全培养基:98%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethanol、10μM SB431542、0.25μM LDN193189和100μM维生素A酸、25μg/ml胰岛素。
N2培养基:97%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethano、1%N2 supplement、1μM SAG和100μM维生素A酸。
B27培养基:95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethanol、1%N2 supplement、2%B27supplement和1μM SAG和100μM维生素A酸、25μg/ml胰岛素。
OPC成熟培养基-不含葛根素:95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethanol、1%N2 supplement、2%B27supplement、10ng/mLPDGF-AA、10ng/mL IGF-1、5ng/mL HGF、10ng/mL NT3、60ng/mL T3、100ng/mL Biotin、1μMcAMP、25μg/ml胰岛素。
OPC成熟培养基-含葛根素:95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM2-Mercaptoethanol、1%N2 supplement、2%B27supplement、10ng/mLPDGF-AA、10ng/mL IGF-1、5ng/mL HGF、10ng/mL NT3、60ng/mL T3、100ng/mL Biotin、1μMcAMP、25μM Puerarin(葛根素)、25μg/ml胰岛素。
按照以下步骤诱导神经干细胞,并验证是否高效分化出少突胶质细胞:
1.从第0天起,iPSC由E8完全培养基(STEMCELL,05991)更换为神经诱导完全培养基。
2.置于37℃,5%CO2培养箱。
3.之后第1-7天,每天换液。
4.每天仔细观察细胞形态改变。
5.从第8天起,神经诱导完全培养基更换为N2培养基。
6.置于37℃,5%CO2培养箱。
7.之后第8-11天,每天换液。
8.每天仔细观察细胞形态改变。
9.从第12天起,将N2培养基更换为B27培养基,细胞由贴壁培养转换为悬浮培养。
10.在第12天,将旧的培养基吸掉,每孔加入B27培养基;
11.用灭菌的刀片刮细胞,至少刮20下,然后分别将孔旋转90°和45°,同样各刮至少20下;
12.用细胞刮将整个孔沿着刮线将细胞刮下;
13.用1ml枪头轻轻吹打3-5次,然后将1孔细胞转移到低吸附的6孔板的两个孔中。然后每个孔分别补加B27培养基,使每孔的终体积为3ml;置于37℃,5%CO2培养箱。
14.之后第12-19天,隔天换一次液。
15.每天仔细观察细胞形态改变。
16.从第20天起,将B27培养基更换为OPC成熟培养基-不含葛根素,细胞由贴壁培养转换为悬浮培养。
17.在第20天,用1ml枪头将球形聚集体转移到15ml离心管中,静置3min使球形聚集体沉到离心管底部,吸掉2/3的旧的培养基,然后重新补加同样体积的OPC成熟培养基,然后将球形聚集体重新转回到原来的低吸附6孔板中。
18.之后第20-30天,隔天换一次液。
在第16天按通用方法进行免疫荧光检测,结果如图1所示:
A:细胞中同时存在Olig2+(绿色,OPC marker)的细胞和NKX2.2+(红色,神经干细胞marker)的细胞,但是没有同时表达Olig2+(绿色)和NKX2.2+(红色)的细胞,说明还没有成熟的OPC细胞;
B:细胞中同时存在Olig2+(绿色,OPC marker)的细胞和Nestin+(红色,神经干细胞marker)阳性的细胞,但是没有同时表达Olig2+(绿色)和Nestin+(红色)的细胞,说明还没有成熟的OPC细胞;
C:细胞中同时存在Olig2+(绿色,OPC marker)的细胞和PAX6+(红色,神经干细胞marker)的细胞,但是没有同时表达Olig2+(绿色)和PAX6+(红色)的细胞,说明还没有成熟的OPC细胞。
在第33天按通用方法进行免疫荧光检测;结果如图2所示诱导day33之后Olig2和Nkx2.2共表达于细胞中,表明诱导OPC取得成功。
实施例2、OPC成熟培养基中葛根素对诱导少突胶质细胞的影响
按照实施例1的方法配置神经诱导完全培养基、N2培养基、B27培养基、OPC成熟培养基;并且配置不含有葛根素的OPC成熟培养基;以含有葛根素或不含有葛根素的OPC成熟培养基为对照,探索葛根素在诱导少突胶质细胞形成中的影响。
在培养的第37天,定量统计少突胶质细胞占全部细胞的百分比,统计如图3所示,进行免疫荧光分析,结果如图4所示。
使用不含葛根素的OPC成熟培养基的为对照组,对照组能够产生约21%少突胶质细胞,添加葛根素后可以产生约36%少突胶质细胞。证明OPC成熟培养基中添加葛根素有利于少突胶质细胞的产生。
Claims (10)
1.一种神经诱导完全培养基,所述神经诱导完全培养基包含第一小分子化合物组合,所述第一小分子化合物组合包括胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂中的至少一种;
优选地,所述胰岛素的工作浓度为25μg/ml;
优选地,所述TGF-β信号通路抑制剂包括SB431542、Dorsomorphin、A83-01、LDN193189、RepSox、SB 525334、DMH-1、SB-505124、BMP signaling agonist sb4、GW788388、ITD-1、LSKL,SD-208、LDN-212854、K02288、LDN-214117、R-268712、SM16、A 77-01、ALK2-IN-2、PD-161570、pm26、TGF-β1peptide TFA、Isosaponarin、BIO-013077-01;
优选地,所述TGF-β信号通路抑制剂选用SB431542;
优选地,所述SB431542的工作浓度为10μM;
优选地,所述BMP信号通路抑制剂包括Dorsomorphin、A83-01、LDN193189、RepSox、SB525334、DMH-1、SB-505124、BMP signaling agonist sb4、GW788388、ITD-1、LSKL、SD-208、LDN-212854、K02288、LDN-214117、R-268712、SM16、A 77-01、ALK2-IN-2、PD-161570、pm26、TGF-β1peptide TFA、Isosaponarin、BIO-013077-01;
优选地,所述BMP信号通路抑制剂是LDN193189;
优选地,所述LDN193189的工作浓度为0.25μM;
优选地,所述RAR核受体的激动剂包括维生素A酸、Rapamycin、3-Methyladenine、Acetylcysteine、5-Fluorouracil,Hydrocortisone、Docetaxel、Rosiglitazone、Estradiol、Melatonin、GW9662、Nicotinamide、Cytarabine、Isoprenalinehydrochloride、Prostaglandin E2、Acetaminophen、(-)-Epigallocatechin Gallate、β-Nicotinamide mononucleotide、Calcitriol、DHEA、Liothyronine、NAD+、Luteolin、Thymidine、Docosahexaenoic Acid、Kaempferol、Palmitic acid、Cyclopamine、Genistein、L-Glutathione reduced;
优选地,所述RAR核受体的激动剂是维生素A酸;
优选地,所述维生素A酸的工作浓度为100μM;
优选地,所述神经诱导完全培养基是由第一基础培养基、非必须氨基酸、谷氨酰胺、还原剂和第一小分子化合物组合组成;
优选地,所述第一基础培养基包括TeSR-E8、mTESR1、E8、Essential 8TMMedium、达尔伯克改良伊格尔培养基、最小必需培养基、伊格尔基本培养基、F-10、F-12、α-最小必需培养基、G-最小必需培养基、IMPM、AmnioMax、新型二代羊水培养基、Chang’s培养基、MesemCult-XF培养基、RPMI1640、Ham’s F12、DMEM/F12、Ham's F-12K Medium、Hepato ZYME-SFM、William’s E Medium、Waymouth’s Medium或Hepatocyte Culture Medium;
优选地,所述第一基础培养基是DMEM/F12;
优选地,所述非必须氨基酸包括丙氨酸、精氨酸、天门冬氨酸、胱氨酸、脯氨酸、酪氨酸;
优选地,所述谷氨酰胺替换为GlutaMAX-I;
优选地,所述还原剂包括但不限于β-巯基乙醇、二硫苏糖醇、二硫赤藓糖醇、还原型谷胱甘肽、半胱氨酸、硫代胺甲酸盐、二硫亚磺酸钠、抗坏血酸盐、二氯化锡或硼氢化钠;
优选地,所述还原剂是β-巯基乙醇;
优选地,所述神经诱导完全培养基是由98%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethanol、10μM SB431542、0.25μM LDN193189和100μM维生素A酸、25μg/ml胰岛素组成。
2.一种N2培养基,所述N2培养基包含第二小分子化合物组合;
优选地,所述第二小分子化合物组合包括RAR核受体的激动剂和/或Hedgehog信号通路激活剂;
优选地,所述RAR核受体的激动剂包括维生素A酸、Rapamycin、3-Methyladenine、Acetylcysteine、5-Fluorouracil,Hydrocortisone、Docetaxel、Rosiglitazone、Estradiol、Melatonin、GW9662、Nicotinamide、Cytarabine、Isoprenalinehydrochloride、Prostaglandin E2、Acetaminophen、(-)-Epigallocatechin Gallate、β-Nicotinamide mononucleotide、Calcitriol、DHEA、Liothyronine、NAD+、Luteolin、Thymidine、Docosahexaenoic Acid、Kaempferol、Palmitic acid、Cyclopamine、Genistein、L-Glutathione reduced;
优选地,所述RAR核受体的激动剂是维生素A酸;
优选地,所述维生素A酸的工作浓度为100μM;
优选地,所述Hedgehog信号通路激活剂包括SAG、Cyclopamine、Purmorphamine、20(S)-Hydroxycholesterol、Halcinonide、Jervine、ALLO-2、IHR-Cy3;
优选地,所述Hedgehog信号通路激活剂是SAG;
优选地,所述SAG的工作浓度为1μM;
优选地,所述N2培养基是由第二基础培养基、非必须氨基酸、谷氨酰胺、还原剂、N2supplement和第二小分子化合物组合组成;
优选地,所述第二基础培养基包括TeSR-E8、mTESR1、E8、Essential 8TMMedium、达尔伯克改良伊格尔培养基、最小必需培养基、伊格尔基本培养基、F-10、F-12、α-最小必需培养基、G-最小必需培养基、IMPM、AmnioMax、新型二代羊水培养基、Chang’s培养基、MesemCult-XF培养基、RPMI1640、Ham’s F12、DMEM/F12、Ham's F-12K Medium、Hepato ZYME-SFM、William’s E Medium、Waymouth’s Medium或Hepatocyte Culture Medium;
优选地,所述第二基础培养基是DMEM/F12;
优选地,所述非必须氨基酸包括丙氨酸、精氨酸、天门冬氨酸、胱氨酸、脯氨酸、酪氨酸;
优选地,所述谷氨酰胺替换为GlutaMAX-I;
优选地,所述还原剂包括但不限于β-巯基乙醇、二硫苏糖醇、二硫赤藓糖醇、还原型谷胱甘肽、半胱氨酸、硫代胺甲酸盐、二硫亚磺酸钠、抗坏血酸盐、二氯化锡或硼氢化钠;
优选地,所述N2 supplement是商品化的产品或自行配制获得;
优选地,所述还原剂是β-巯基乙醇;
优选地,所述N2培养基是由97%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethano、1%N2 supplement、1μM SAG和100μM维生素A酸组成。
3.一种B27培养基,所述B27培养基包含第三小分子化合物组合;
优选地,所述第三小分子化合物组合包括胰岛素、RAR核受体的激动剂和Hedgehog信号通路激活剂中的至少一种;
优选地,所述胰岛素的工作浓度为25μg/ml;
优选地,所述RAR核受体的激动剂包括维生素A酸、Rapamycin、3-Methyladenine、Acetylcysteine、5-Fluorouracil,Hydrocortisone、Docetaxel、Rosiglitazone、Estradiol、Melatonin、GW9662、Nicotinamide、Cytarabine、Isoprenalinehydrochloride、Prostaglandin E2、Acetaminophen、(-)-Epigallocatechin Gallate、β-Nicotinamide mononucleotide、Calcitriol、DHEA、Liothyronine、NAD+、Luteolin、Thymidine、Docosahexaenoic Acid、Kaempferol、Palmitic acid、Cyclopamine、Genistein、L-Glutathione reduced;
优选地,所述RAR核受体的激动剂是维生素A酸;
优选地,所述维生素A酸的工作浓度为100μM;
优选地,所述Hedgehog信号通路激活剂包括SAG、Cyclopamine、Purmorphamine、20(S)-Hydroxycholesterol、Halcinonide、Jervine、ALLO-2、IHR-Cy3;
优选地,所述Hedgehog信号通路激活剂是SAG;
优选地,所述SAG的工作浓度为1μM;
优选地,所述B27培养基是由第三基础培养基、非必须氨基酸、谷氨酰胺、还原剂、N2supplement、B27 supplement和第三小分子化合物组合组成;
优选地,所述第三基础培养基包括TeSR-E8、mTESR1、E8、Essential 8TMMedium、达尔伯克改良伊格尔培养基、最小必需培养基、伊格尔基本培养基、F-10、F-12、α-最小必需培养基、G-最小必需培养基、IMPM、AmnioMax、新型二代羊水培养基、Chang’s培养基、MesemCult-XF培养基、RPMI1640、Ham’s F12、DMEM/F12、Ham's F-12K Medium、Hepato ZYME-SFM、William’s E Medium、Waymouth’s Medium或Hepatocyte Culture Medium;
优选地,所述第三基础培养基是DMEM/F12;
优选地,所述非必须氨基酸包括丙氨酸、精氨酸、天门冬氨酸、胱氨酸、脯氨酸、酪氨酸;
优选地,所述谷氨酰胺替换为GlutaMAX-I;
优选地,所述还原剂包括但不限于β-巯基乙醇、二硫苏糖醇、二硫赤藓糖醇、还原型谷胱甘肽、半胱氨酸、硫代胺甲酸盐、二硫亚磺酸钠、抗坏血酸盐、二氯化锡或硼氢化钠;
优选地,所述N2 supplement是商品化的产品或自行配制获得;
优选地,所述B27 supplement是商品化的产品或自行配制获得;
优选地,所述B27培养基是由95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM 2-Mercaptoethanol、1%N2 supplement、2%B27supplement和1μM SAG和100μM维生素A酸、25μg/ml胰岛素组成。
4.一种OPC成熟培养基,所述OPC成熟培养基中包括第四小分子化合物组合;
优选地,所述第四小分子化合物组合中含有5-HT2C受体拮抗剂;
优选地,所述5-HT2C受体拮抗剂包括Puerarin、Harmine、SCH-23390hydrochloride、Olanzapine、GTS-21dihydrochloride、Thioridazine hydrochloride、Risperidone、SB-269970hydrochloride、Dihydroergotamine mesylate、Amitriptyline hydrochloride、Aripiprazole、Ketanserin、WAY-100635Maleate、Buspirone hydrochloride、Cisapride、Methiothepin mesylate、Trazodone hydrochloride、Pindolol、Quetiapine、Lumateperone tosylate、Perphenazine、8-OH-DPAT、Levomepromazine、SB-224289hydrochloride、Volinanserin、Ziprasidone、Paliperidone、Sertindole、Alprenololhydrochloride、Asenapine maleate;
优选地,所述5-HT2C受体拮抗剂是Puerarin;
优选地,所述Puerarin的工作浓度为25μM;
优选地,所述第四小分子化合物组合中还含有胰岛素、PDGF-AA、IGF-1、HGF、NT3、T3、Biotin、cAMP;
优选地,所述第四小分子化合物组合中还含有25μg/ml胰岛素、10ng/mL PDGF-AA、10ng/mL IGF-1、5ng/mL HGF、10ng/mL NT3、60ng/mL T3、100ng/mL Biotin、1μM cAMP;
优选地,所述OPC成熟培养基由第四基础培养基、非必须氨基酸、谷氨酰胺、还原剂、N2supplement、B27 supplement至少一种组成;
优选地,所述第四基础培养基包括TeSR-E8、mTESR1、E8、Essential 8TMMedium、达尔伯克改良伊格尔培养基、最小必需培养基、伊格尔基本培养基、F-10、F-12、α-最小必需培养基、G-最小必需培养基、IMPM、AmnioMax、新型二代羊水培养基、Chang’s培养基、MesemCult-XF培养基、RPMI1640、Ham’s F12、DMEM/F12、Ham's F-12K Medium、Hepato ZYME-SFM、William’s E Medium、Waymouth’s Medium或Hepatocyte Culture Medium;
优选地,所述第四基础培养基是DMEM/F12;
优选地,所述还原剂包括但不限于β-巯基乙醇、二硫苏糖醇、二硫赤藓糖醇、还原型谷胱甘肽、半胱氨酸、硫代胺甲酸盐、二硫亚磺酸钠、抗坏血酸盐、二氯化锡或硼氢化钠;
优选地,所述非必须氨基酸包括丙氨酸、精氨酸、天门冬氨酸、胱氨酸、脯氨酸、酪氨酸;
优选地,所述N2 supplement是商品化的产品或自行配制获得;
优选地,所述B27 supplement是商品化的产品或自行配制获得;
优选地,所述谷氨酰胺替换为GlutaMAX-I;
优选地,所述OPC成熟培养基的组成选自以下任意一种:
1)95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM2-Mercaptoethanol、1%N2 supplement、2%B27supplement、10ng/mL PDGF-AA、10ng/mLIGF-1、5ng/mL HGF、10ng/mL NT3、60ng/mL T3、100ng/mL Biotin、1μM cAMP、25μMPuerarin、25μg/ml胰岛素;
2)95%DMEM/F-12培养基、1%非必须氨基酸、1%GlutaMAX-I、0.1mM2-Mercaptoethanol、1%N2 supplement、2%B27supplement、10ng/mL PDGF-AA、10ng/mLIGF-1、5ng/mL HGF、10ng/mL NT3、60ng/mL T3、100ng/mL Biotin、1μM cAMP、25μg/ml胰岛素。
5.一种诱导神经干细胞的方法,所述方法包括使用权利要求1所述神经诱导完全培养基和/或权利要求2所述N2培养基进行细胞培养;
优选地,所述方法包括使用权利要求1所述神经诱导完全培养基和权利要求2所述N2培养基进行细胞培养。
6.一种诱导少突胶质细胞的方法,所述方法包括使用权利要求1所述神经诱导完全培养基、权利要求2所述N2培养基、权利要求3所述B27培养基、权利要求4所述OPC成熟培养基中至少一种培养基进行细胞培养;
优选地,所述方法是依次使用权利要求1所述神经诱导完全培养基、权利要求2所述N2培养基、权利要求3所述B27培养基、权利要求4所述OPC成熟培养基进行细胞培养。
7.如权利要求5或6所述的方法,其特征在于,所述细胞培养的起始细胞是干细胞,所述干细胞包括ESC、诱导性多能干细胞、拟胚体、细胞造血干细胞、神经干细胞、间充质干细胞、皮肤干细胞、脂肪干细胞、脐血干细胞中的一种或多种;
优选地,所述干细胞是诱导性多能干细胞。
8.一种细胞群,其特征在于,所述细胞群选自以下一种:
1)由权利要求5所述方法制备得到的细胞群,其特征在于,所述细胞群中包括表达NKX2.2+、Nestin+、PAX6+中至少一种的细胞;
优选地,所述细胞群中存在表达Olig2+的细胞;
2)由权利要求6所述方法制备得到的细胞群,其特征在于,所述细胞群中少突胶质细胞占全部细胞比例的至少20%;优选地,至少21%、25%、30%、35%、36%;
优选地,所述少突胶质细胞是表达Olig2、Nkx2.2、O4和MBP中至少一种的细胞;
优选地,所述少突胶质细胞是Olig2和Nkx2.2共表达的细胞;或者,所述少突胶质细胞是O4和MBP共表达的细胞。
9.一种诱导少突胶质细胞的试剂盒,所述试剂盒中包括胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂、Hedgehog信号通路激活剂、5-HT2C受体拮抗剂中的一种或多种;
优选地,所述TGF-β信号通路抑制剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂;
优选地,所述BMP信号通路抑制剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂;
优选地,所述RAR核受体的激动剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂或权利要求2所述的N2培养基使用的TGF-β信号通路抑制剂或权利要求3所述的B27培养基使用的TGF-β信号通路抑制剂;
优选地,所述Hedgehog信号通路激活剂是如权利要求1所述的神经诱导完全培养基使用的Hedgehog信号通路激活剂或权利要求2所述的N2培养基使用的Hedgehog信号通路激活剂或权利要求3所述的B27培养基使用的Hedgehog信号通路激活剂;
优选地,所述5-HT2C受体拮抗剂是如权利要求4所述的OPC成熟培养基使用的Hedgehog信号通路激活剂;
优选地,所述试剂盒中包括配制权利要求1所述的神经诱导完全培养基、权利要求2所述的N2培养基、权利要求3所述的B27培养基、权利要求4所述的OPC成熟培养基所需要的试剂。
10.一种应用,其特征在于,所述应用选自以下任意一种:
1)权利要求9所述的试剂盒、权利要求1所述的神经诱导完全培养基、权利要求2所述的N2培养基、权利要求3所述的B27培养基、权利要求4所述的OPC成熟培养基、胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂、Hedgehog信号通路激活剂、5-HT2C受体拮抗剂、包含5-HT2C受体拮抗剂的小分子化合物组合任意一种在诱导少突胶质细胞中的应用;
优选地,所述TGF-β信号通路抑制剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂;
优选地,所述BMP信号通路抑制剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂;
优选地,所述RAR核受体的激动剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂或权利要求2所述的N2培养基使用的TGF-β信号通路抑制剂或权利要求3所述的B27培养基使用的TGF-β信号通路抑制剂;
优选地,所述Hedgehog信号通路激活剂是如权利要求1所述的神经诱导完全培养基使用的Hedgehog信号通路激活剂或权利要求2所述的N2培养基使用的Hedgehog信号通路激活剂或权利要求3所述的B27培养基使用的Hedgehog信号通路激活剂;
优选地,所述5-HT2C受体拮抗剂是如权利要求4所述的OPC成熟培养基使用的Hedgehog信号通路激活剂;
优选地,所述包含5-HT2C受体拮抗剂的小分子化合物组合是权利要求4所述的OPC成熟培养基使用的第四小分子化合物组合;
2)由权利要求6所述的方法制备得到的细胞在治疗中枢神经系统疾病中的用途;
优选地,所述中枢神经系统疾病包括但不限于神经退行性疾病、脱髓鞘疾病、癫痫、脑外伤、休克、痴呆、青光眼、多发性硬化症、神经系统损伤后再生以及精神类疾病;
优选地,所述脱髓鞘疾病包括脑卒中、多发性硬化、视神经脊髓炎、急性播散性脑脊髓炎、弥漫性硬化、同心圆硬化、脑白质营养不良、脑桥中央髓鞘溶解症、急性炎症性脱髓鞘性多发性神经病、慢性炎症性脱髓鞘性多发性神经病、缺血-缺氧性病引起的白质脑病、营养缺乏性疾病引起的亚急性联合变性、病毒感染引起的亚急性硬化性全脑炎或进行性多灶性白质脑病、糖尿病性神经病、系统性红斑狼疮的神经病变、肾上腺脑白质营养不良、白质消融性疾病、佩利措伊斯-梅茨巴赫病;
3)权利要求9所述的试剂盒、权利要求1所述的神经诱导完全培养基、权利要求2所述的N2培养基、胰岛素、TGF-β信号通路抑制剂、BMP信号通路抑制剂、RAR核受体的激动剂、Hedgehog信号通路激活剂任意一种在诱导神经干细胞中的应用;
优选地,所述TGF-β信号通路抑制剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂;
优选地,所述BMP信号通路抑制剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂;
优选地,所述RAR核受体的激动剂是如权利要求1所述的神经诱导完全培养基使用的TGF-β信号通路抑制剂或权利要求2所述的N2培养基使用的TGF-β信号通路抑制剂或权利要求3所述的B27培养基使用的TGF-β信号通路抑制剂;
优选地,所述Hedgehog信号通路激活剂是如权利要求1所述的神经诱导完全培养基使用的Hedgehog信号通路激活剂或权利要求2所述的N2培养基使用的Hedgehog信号通路激活剂或权利要求3所述的B27培养基使用的Hedgehog信号通路激活剂。
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114262686A (zh) * | 2022-03-02 | 2022-04-01 | 深圳市夏同生物医药科技有限公司 | 一种少突胶质细胞的制备方法及应用 |
CN115612667A (zh) * | 2022-11-02 | 2023-01-17 | 广州瑞臻再生医学科技有限公司 | 一种诱导多能干细胞来源的前脑少突胶质细胞的制备方法 |
WO2023010897A1 (zh) * | 2021-08-05 | 2023-02-09 | 呈诺再生医学科技(珠海横琴新区)有限公司 | 人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用 |
WO2023241715A1 (zh) * | 2022-06-16 | 2023-12-21 | 中国科学院动物研究所 | 视黄酸受体激活剂及其组合在哺乳动物再生修复中的应用 |
Families Citing this family (1)
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---|---|---|---|---|
US20240043798A1 (en) * | 2022-08-08 | 2024-02-08 | Trailhead Biosystems Inc. | Methods and compositions for generating oligodendrocyte progenitor cells |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20150352154A1 (en) * | 2013-02-06 | 2015-12-10 | University Of Rochester | Induced pluripotent cell-derived oligodendrocyte progenitor cells for the treatment of myelin disorders |
CN106456672A (zh) * | 2014-05-22 | 2017-02-22 | 纽约干细胞基金会有限公司 | 源于多能干细胞的功能性少突胶质细胞及其制备和使用方法 |
CN107109367A (zh) * | 2014-10-24 | 2017-08-29 | 大日本住友制药株式会社 | 神经组织的制备方法 |
CN107208045A (zh) * | 2014-11-25 | 2017-09-26 | 宾州研究基金会 | 人神经胶质细胞转化成用于大脑和脊髓修复的神经细胞的化学重编程 |
CN108384755A (zh) * | 2018-02-08 | 2018-08-10 | 北京呈诺医学科技有限公司 | 一种高效、快捷的诱导性多能干细胞向神经干细胞分化的方法 |
CN108624560A (zh) * | 2018-06-01 | 2018-10-09 | 南京艾尔普再生医学科技有限公司 | 一种分化培养基及少突胶质前体细胞的制备方法 |
CN112469818A (zh) * | 2018-04-17 | 2021-03-09 | 凯斯西储大学 | 人皮层球体中髓鞘少突胶质细胞的诱导 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2019241620A1 (en) * | 2018-06-14 | 2019-12-19 | Academia Sinica | Method to generate induced oligodendrocyte-lineage cells and treatment using such cells |
CN113564122B (zh) * | 2021-08-05 | 2022-04-08 | 呈诺再生医学科技(珠海横琴新区)有限公司 | 人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用 |
-
2021
- 2021-08-05 CN CN202110894940.XA patent/CN113564122B/zh active Active
-
2022
- 2022-04-22 WO PCT/CN2022/088517 patent/WO2023010897A1/zh unknown
-
2023
- 2023-11-28 US US18/521,956 patent/US20240093148A1/en active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20150352154A1 (en) * | 2013-02-06 | 2015-12-10 | University Of Rochester | Induced pluripotent cell-derived oligodendrocyte progenitor cells for the treatment of myelin disorders |
CN106456672A (zh) * | 2014-05-22 | 2017-02-22 | 纽约干细胞基金会有限公司 | 源于多能干细胞的功能性少突胶质细胞及其制备和使用方法 |
CN107109367A (zh) * | 2014-10-24 | 2017-08-29 | 大日本住友制药株式会社 | 神经组织的制备方法 |
CN107208045A (zh) * | 2014-11-25 | 2017-09-26 | 宾州研究基金会 | 人神经胶质细胞转化成用于大脑和脊髓修复的神经细胞的化学重编程 |
CN108384755A (zh) * | 2018-02-08 | 2018-08-10 | 北京呈诺医学科技有限公司 | 一种高效、快捷的诱导性多能干细胞向神经干细胞分化的方法 |
CN112469818A (zh) * | 2018-04-17 | 2021-03-09 | 凯斯西储大学 | 人皮层球体中髓鞘少突胶质细胞的诱导 |
CN108624560A (zh) * | 2018-06-01 | 2018-10-09 | 南京艾尔普再生医学科技有限公司 | 一种分化培养基及少突胶质前体细胞的制备方法 |
Non-Patent Citations (2)
Title |
---|
GARY X. D’SOUZA 等: "The application of in vitro-derived human neurons in neurodegenerative disease modeling", 《J NEUROSCI RES.》 * |
SAMANTHA N.等: "Growing Glia:Cultivating Human Stem Cell Models of Gliogensis in Health and Disease", 《FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY 》 * |
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2023010897A1 (zh) * | 2021-08-05 | 2023-02-09 | 呈诺再生医学科技(珠海横琴新区)有限公司 | 人诱导性多能干细胞向少突胶质细胞分化的方法,试剂盒以及应用 |
CN114262686A (zh) * | 2022-03-02 | 2022-04-01 | 深圳市夏同生物医药科技有限公司 | 一种少突胶质细胞的制备方法及应用 |
WO2023165062A1 (zh) * | 2022-03-02 | 2023-09-07 | 深圳市夏同生物医药科技有限公司 | 一种少突胶质细胞的制备方法及应用 |
WO2023241715A1 (zh) * | 2022-06-16 | 2023-12-21 | 中国科学院动物研究所 | 视黄酸受体激活剂及其组合在哺乳动物再生修复中的应用 |
CN115612667A (zh) * | 2022-11-02 | 2023-01-17 | 广州瑞臻再生医学科技有限公司 | 一种诱导多能干细胞来源的前脑少突胶质细胞的制备方法 |
CN115612667B (zh) * | 2022-11-02 | 2023-09-08 | 广州瑞臻再生医学科技有限公司 | 一种诱导多能干细胞来源的前脑少突胶质细胞的制备方法 |
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