CN116751735A - 一种脐带间充质干细胞的无血清培养方法 - Google Patents
一种脐带间充质干细胞的无血清培养方法 Download PDFInfo
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Abstract
本发明涉及一种脐带间充质干细胞的无血清培养方法,所述方法的培养基中特异性的添加特异性诱导Pdx1高表达的多肽。所述的方法能够特异性的提高脐带血间充质干细胞的培养密度的同时,还能够有效的促进脐带血间充质干细胞诱导分化为胰岛样细胞,从而有效的用于糖尿病的治疗。
Description
技术领域
本申请涉及生物领域,具体的涉及一种脐带间充质干细胞的无血清培养方法。
背景技术
干细胞以其极强的自我更新能力及多向分化潜能,目前备受研究人员关注,已经被广泛应用到血液系统、神经系统、循环系统和免疫系统等多个系统疾病的治疗中,并取得了初步疗效。在内分泌系统疾病的治疗中,干细胞也开始得到广泛应用,尤其是在糖尿病的治疗中。已经逐渐成为研究人员寻找胰岛细胞替代物的新资源。
血中除富含造血干细胞外,研究人员还发现了不同表型特征的其他细胞类型,这些细胞除具有支持造血功能外,还可向诸如软骨、骨、脂肪和肌细胞等多种类型的中胚层细胞分化,提示脐带血中存在MSCS的成分。此后,随着有关脐血细胞研究的深入,研究者的结论也逐渐趋于一致,即脐带血中的确存在MSCs,而且证实,其形态学和生物学特性与骨髓来源的MSCS相似,因此可以考虑作为细胞治疗和基因治疗的载体细胞。但面临的问题是,脐带血中所含的MSCs数量极少,每100万个脐带血单个核细胞中仅含0.05-2.8个MSCs。这就导致脐带血MSCs的培养成功率很低,一般报道只有20%-30%,如此低的培养成功率极大浪费了脐带血资源。在脐带血样本采集和分离条件控制的基础上,培养成功的关键在于每次采集样本的体积不少于40ml,并在12小时内尽快完成分离培养,分离的单个核细胞的数量要大于1×108,并要以1×106/cm2密度接种于胎牛血清包被的培养器皿中。研究报道,除了细胞的接种密度外,适宜的细胞因子种类和用量也是MSCs培养成功的关键。在传统的培养方法基础上,通过添加细胞因子IL-3和GM-CSF可以使脐带血MSCs的原代培养成功率由传统方法的25%提高到90%以上。
糖尿病是一种由于胰岛素分泌缺陷或胰岛素作用障碍所致的以高血糖为特征的代谢性疾病。据国际糖尿病联合会报道:全世界大约有3亿人患有糖尿病,并呈上升趋势。1型糖尿病多由胰岛β细胞自身免疫破坏所致,而较为常见的2型糖尿病主因胰岛素抵抗和胰岛β细胞功能障碍所致。对于糖尿病患者,尤其是1型糖尿病患者,必须使用外源性胰岛素来控制血糖稳定。尽管胰岛素治疗效果不错,但其不能消除糖尿病所带来的慢性并发症,例如心血管疾病、肾功能衰竭、视网膜病变和神经病变等。将正常鼠胰岛用1型寡聚胶原包裹移植到糖尿病鼠皮下,发现其可以降低血糖并维持血糖稳定90d。但由于胰岛供者的缺少也限制了此种方法在临床上的应用。近些年来,科学家们发现移植间充质干细胞可以治疗糖尿病。间充质干细胞是一类具有多向分化潜能的多能干细胞,可诱导分化成骨细胞、脂肪细胞、心肌细胞、胰岛β细胞。将人脐带间充质干细胞移植到1型糖尿病大鼠体内,可降低其血糖并修复胰岛功能;将脂肪间充质干细胞移植到2型糖尿病大鼠体内,不仅可以降低血糖而且可改善相应并发症。目前认为间充质干细胞治疗糖尿病主要机制为:①在胰腺微环境下定向诱导分化为胰岛β细胞,代替受损胰岛功能;②通过释放抗炎和免疫调节因子,修复胰岛功能;③通过归巢和旁分泌多种细胞因子以及核酸片段,改善胰岛细胞微环境。为了更好地发挥间充质干细胞治疗糖尿病的效果,科学家们对其进行体外诱导,在相关因子的作用下使其分化为能分泌胰岛素的细胞,即胰岛样细胞,将其移植到糖尿病鼠体内,发现其降糖效果优于直接移植间充质干细胞。
但是目前,间充质干细胞诱导分化为胰岛样细胞的效率不高还有待进一步的提高。因此,为了治疗糖尿病,首选需要提高干细胞的诱导效率。无论是进行胰岛移植还是干细胞移植,体内实验结果表明移植大量的胰岛或干细胞才能达到良好的血糖控制目标。然而目前的国内外研究虽然发现了多种干细胞可以分化为胰岛样细胞,但是分化来的胰岛样细胞数目占分化前干细胞数目的比例很低,胰岛样细胞较正常细胞胰岛素分泌量少,因此要获得大量有良好分泌功能的胰岛样细胞是继续解决的技术问题。探索干细胞向胰岛样细胞的分化机制、优化干细胞诱导分化条件以及提高干细胞向胰岛样细胞的转化效率等仍是目前亟待解决的问题。
发明内容
本发明克服现有技术的缺陷,提供了一种改进的脐带血间充质干细胞培养基及其培养方法。
所述的方法能够特异性的提高脐带血间充质干细胞的培养密度的同时,还能够有效的促进脐带血间充质干细胞诱导分化为胰岛样细胞,从而有效的用于糖尿病的治疗。
具体的,本发明研究发现,胰十二指肠同源框因子-1,Pdx1基因参与了胰岛细胞的分化成熟过程。Pdx1基因是胰腺发生和胰岛发育的一个重要调节基因。现有技术中,通过转基因Pdx1到干细胞中进行了胰岛样细胞的诱导和分化,但是该基因的过表达一方面影响细胞本身的代谢平衡,另外一方面,转基因的操作本身复杂并且基因表达不稳定影响细胞分化的稳定性及持续性。在此基础上,本发明通过筛选鉴定,获得了能够特异性诱导Pdx1高表达的多肽,所述多肽是通过计算机模拟结合筛选并通过特异性鉴定获得的。
具体的,所述的特异性诱导Pdx1高表达的多肽其氨基酸序列如SEQ ID NO:1所示。
进一步的,本发明提供一种培养脐带血间充质干细胞的无血清培养基,所述无血清培养基包括基础培养基以及特异性诱导Pdx1高表达的多肽其氨基酸序列如SEQ ID NO:1所示。
具体的,所述的基础培养基为StemCell MesenCultTM间充质干细胞培养基,或者其他的无血清培养基。
如本文所用的,无血清培养基,可以在基础培养基的基础上添加其他合适的组分。术语“基础培养基”是指可有效支持培养细胞生长的氨基酸、维生素、盐和营养素的溶液,尽管通常地,这些化合物并不支持细胞生长除非添加了其它化合物。营养素包括可被细胞代谢的碳源(例如糖如葡萄糖),以及细胞存活必需的其它化合物。存在细胞自身不能合成的化合物,这是由于缺乏编码合成该化合物(例如必需氨基酸)所必需蛋白的一个或多个基因,或与细胞可以合成的化合物相关,因为它们的特定的发育状态,编码必需生物合成蛋白的基因不按足够水平表达。许多基础培养基是哺乳动物细胞培养领域所已知的,例如达尔贝科改良依格培养基(DMEM),敲除-DMEM(KO-DMEM),和DMEM/F12,尽管可采用在基本上不分化状态下支持灵长类胚胎干细胞生长的任何基础培养基。
本领域技术人员应理解,用于增殖细胞的细胞培养基可影响一种或多种细胞特征,包括但不限于:非致瘤性、非致癌性、以贴壁细胞形式生长、以非贴壁细胞形式生长、具有上皮样形态学特征、在培养时支持各种病毒复制和支持流感病毒复制达到本文所述高效价。为降低被外来物质(例如,支原体、病毒和朊病毒)污染的风险,应最大程度减少或甚至去除在组织培养应用中将血清或动物提取物用于生产治疗(例如疫苗)材料。此外,最大程度减少细胞培养过程中所需操作数量可显著降低污染的可能性。因此,本发明提供可用于采用分批培养方法增殖干细胞的无血清培养基。具体而言,本发明的无血清培养基(本文也称为“本发明的无血清培养基”和“本发明的培养基”)可用于不采用培养基交换或补充的分批培养方法。因此,开发和使用本发明的无血清培养基可克服干细胞生长不好的缺陷。
在另一个实施方式中,本发明的无血清培养基用一种或多种选自下组的培养基组分强化:腐胺、氨基酸、维生素、脂肪酸和核苷。在具体实施方式中,本发明的无血清培养基用一种或多种培养基组分强化,使所述培养基组分的浓度比在常规用于增殖细胞的培养基例如杜尔伯科改良伊格尔培养基/汉氏F12培养基(DMEM/F12)中通常发现的浓度高约1倍、约2倍、约3倍、约4倍或约5倍或更多。
在另一个实施方式中,本发明的无血清培养基用腐胺强化,使腐胺的浓度比在DMEM/F12中通常发现的浓度高约5倍或更多。可强化的脂肪酸包括:不饱和脂肪酸,包括但不限于亚油酸和α-亚麻酸(也称为必需脂肪酸)以及肉豆蔻脑酸、棕榈油酸、油酸、花生四烯酸、二十碳五烯酸、芥酸、二十二碳六烯酸;饱和脂肪酸,包括但不限于丁酸、己酸、辛酸、癸酸、十二烷酸、十四烷酸、十六烷酸、十八烷酸、二十烷酸、二十二烷酸、二十四烷酸;以及含硫脂肪酸,包括硫辛酸。在某些实施方式中,本发明的无血清培养基用除由上述液体补充物提供的脂肪酸以外的额外脂肪酸强化。在一个具体实施方式中,本发明的无血清培养基用亚油酸和亚麻酸强化。在另一个实施方式中,本发明的无血清培养基用亚油酸和亚麻酸强化,使亚油酸和亚麻酸的浓度比在DMEM/F12中通常发现的浓度高约5倍或更多。可强化的氨基酸包括20种标准氨基酸(丙氨酸、精氨酸、天冬酰胺、天冬氨酸、半胱氨酸、谷氨酸、谷氨酰胺、甘氨酸、组氨酸、异亮氨酸、亮氨酸、赖氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、丝氨酸、苏氨酸、色氨酸、酪氨酸和缬氨酸)以及胱氨酸和非标准氨基酸。在某些实施方式中,强化通常称为“必需氨基酸”的非致瘤性MDCK细胞不合成的一种或多种氨基酸。例如,通常将8种氨基酸视作人类必需:苯丙氨酸、缬氨酸、苏氨酸、色氨酸、异亮氨酸、甲硫氨酸、亮氨酸和赖氨酸。在一个具体实施方式中,本发明的无血清培养基用胱氨酸和除谷氨酰胺以外的所有标准氨基酸强化(配制DMEM/F12通常不用谷氨酰胺,将其单独加入),使胱氨酸和标准氨基酸的浓度比在DMEM/F12中通常发现的浓度高约5倍或更多。在某些具体实施方式中,本发明的无血清培养基包含的谷氨酰胺的浓度在约146mg/L到约1022mg/L之间、约292mg/L到约876mg/L之间或约438mg/L到约730mg/L之间。在另一个具体实施方式中,本发明的无血清培养基包含终浓度为584mg/L的谷氨酰胺。
可强化的维生素包括但不限于:抗坏血酸(维生素A)、d-生物素(维生素B7和维生素H)、D-泛酸钙、胆钙化甾醇(维生素D3)、氯化胆碱、氰钴胺素(维生素B12)、麦角钙化固醇(维生素D2)、叶酸(维生素B9)、甲基萘醌类(维生素K2)、肌醇、烟酰胺(维生素B3)、对氨基苯甲酸、泛酸(维生素B5)、叶绿醌(维生素K1)、吡哆醇(维生素B6)、视黄醇(维生素A)、核黄素(维生素B2)、α-生育酚(维生素E)和硫胺素(维生素B1)。在一个具体实施方式中,本发明的无血清培养基用d-生物素、D-泛酸钙、氯化胆碱、氰钴胺素、叶酸、肌醇、烟酰胺、吡哆醇、核黄素和硫胺素进行强化,使所述维生素的浓度比DMEM/F12中通常发现的浓度高约5倍或更多。可强化的核苷包括但不限于:胞苷、尿苷、腺苷、鸟苷、胸苷和次黄嘌呤。在一个具体实施方式中,本发明的无血清培养基用次黄嘌呤和胸苷强化,使次黄嘌呤和胸苷的浓度比在DMEM/F12中通常发现的浓度高约5倍或更多。
可加入细胞培养基的其它组分包括但不限于:碳酸氢钠、碳源(例如葡萄糖)和铁结合试剂。在一个实施方式中,本发明的无血清培养基包含的碳酸氢钠的终浓度在约1200mg/L到约7200mg/L之间、约2400mg/L到约6000mg/L之间或约3600mg/L到约4800mg/L之间。在一个具体实施方式中,本发明的无血清培养基包含终浓度为4400mg/L的碳酸氢钠。在一个实施方式中,本发明的无血清培养基包含葡萄糖作为碳源。在另一个实施方式中,本发明的无血清培养基包含的葡萄糖的终浓度在约1g/L到约10g/L之间、约2g/L到约10g/L之间、约3g/L到约8g/L之间、约4g/L到约6g/L之间或约4.5g/L到约9g/L之间。
进一步的,本发明还提供了一种培养脐带间充质干细胞的方法,所述方法能够诱导脐带间充质干细胞分化为胰岛样细胞,所述方法,包括:将脐带间充质干细胞先在StemCellMesenCultTM间充质干细胞培养基+4nmol/L活化素A+100μg/mL的多肽条件下培养3d,随后在StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+25μg/L表皮生长因子+100μg/mL的多肽培养8d,中间每2的更换新鲜培养液1次;随后StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+10μg/L碱性成纤维细胞生长因子+0.5%胰岛素/转铁蛋白/硒+100μg/mL的多肽诱导培养14d,间隔三天更换新鲜培养液,最后离心收集获得细胞即为诱导分化的胰岛样细胞。
本发明提供一种促进脐带间充质干细胞增殖及分化为胰岛样细胞的培养基,所述培养基包括如下各组:
培养基1:StemCellMesenCultTM间充质干细胞培养基+4nmol/L活化素A+100μg/mL多肽;
培养基2:StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+25μg/L表皮生长因子+100μg/mL多肽;和
培养基3:StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+10μg/L碱性成纤维细胞生长因子+0.5%胰岛素/转铁蛋白/硒+100μg/mL多肽。
有益效果
本发明提供了一种改进的脐带血间充质干细胞培养基及其培养方法,所述的培养基中特异性的添加特异性诱导Pdx1高表达的多肽。所述的方法能够特异性的提高脐带血间充质干细胞的培养密度的同时,还能够有效的促进脐带血间充质干细胞诱导分化为胰岛样细胞,从而有效的用于糖尿病的治疗。
附图说明
图1Pdx-1/GAPDH的相对表达量
图2生长曲线图
具体实施方式
本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。以下实施案例中的方法、设备、材料,如果未进行特别说明,均为本领域常规方法、设备和材料,均可由市场购得。
实施例1脐带间充质干细胞的制备及鉴定
取新生儿出生后娩出的脐带,立即放入80%DMEM培养基加20%双抗(含100U/ml青霉素,100U/ml链霉素)的溶液中,2h内将脐带剪成1mm×1mm×1mm,接种于含有StemCellMesenCultTM间充质干细胞培养基的培养瓶中,将培养瓶倒置于37℃、5%CO2的培养箱中;4h后将培养瓶翻转,24h后添加1ml培养基,3d后全量换干细胞液。之后,每隔3d,全量换液1次。培养5d后,在组织周围会有梭形成纤维样细胞爬出,培养至2周,待细胞融合至80%,去除组织块后传代,第3代细胞即为脐带间充质干细胞。以每管5×106个冻存以便备用。取生长状态良好的第3代细胞,消化并计数,以每管含1×104个细胞,加入PE标记的小鼠抗人的单克隆抗体CD44、CD90、CD34及CD45,以同型对照10μl作为阴性对照,4℃避光孵育30min,磷酸盐缓冲液冲洗后用固定液进行固定,流式仪检测发现,干细胞表面标记CD44和CD90均表达大于95%,而CD34和CD45均小于5%,表明本发明分离制备得到了脐带间充质干细胞。
实施例2特异性诱导Pdx1高表达的多肽功效鉴定
取生长状态良好的实施例1制备的脐带间充质干细胞第3代细胞,消化并计数,以每管含1×104个细胞,接种于含有多肽浓度分别为1、10、50、100、200μg/mL的StemCellMesenCultTM间充质干细胞培养基的培养瓶中,将培养瓶置于37℃、5%CO2的培养箱中3天换液一次,共培养96h;不加多肽的培养基作为对照。
分别提取相应不同浓度的细胞的总蛋白,BCA法进行定量,取100μg蛋白与加样缓冲液混合,100℃变性6min,12%SDS-PAGE胶电泳分离,电转移至硝纤膜,膜在Pdx-1相应抗体中4℃孵育过夜,二抗孵育4h,ECL显色检测。用GAPDH蛋白作内对照,计算Pdx-1/GAPDH的相对表达量。结果如图1所示。
从图1可以看出,在正常的培养基中培养的干细胞中基本不表达Pdx-1蛋白,而本发明的多肽添加到培养基中后,Pdx-1蛋白得到表达,并且具有剂量依赖性的提高干细胞中的PDX-1的表达,在浓度为100和200μg/mL时,Pdx-1/GAPDH的相对表达量分别为2.83和3.02。
实施例3MTT法检测多肽对细胞活性生长状态的影响
取生长状态良好的实施例1制备的脐带间充质干细胞第4代细胞,调整细胞浓度至1×107/L加入96孔板,每孔200μl,放在37℃、5%CO2的培养箱中培养,其中培养基中添加100μg/mL的多肽,不加多肽的StemCell MesenCultTM间充质干细胞培养基作为对照。7d内在特定时间点,取出96孔细胞培养板每孔分别加入20μl MTT(5g/L),37℃放置4h;再每孔加入DMSO 150μl,用酶标仪(波长550nm)测定各孔吸光光度值(D值),以D值代表细胞的相对数量,绘制生长曲线。结果如图2所示。
从图2可以看出,添加了多肽后的培养基与不加多肽的对照培养基相比,在细胞生长特性上显示出更好的促进增殖的效果。这也说明,本发明的诱导Pdx1高表达的多肽除了诱导Pdx1高表达之外,还能促进干细胞的增殖,在7d时,加多肽的OD550值为3.71,而不加肽的OD550值为3.09,促增殖效果明显。
实施例4脐带间充质干细胞诱导分化为胰岛样细胞实验
取生长状态良好的实施例1制备的脐带间充质干细胞第4代细胞,细胞达80%融合后开始诱导。实验分为3组,即细胞因子诱导组,多肽联合细胞因子组,干细胞对照组。
细胞因子诱导组为将干细胞先在StemCellMesenCultTM间充质干细胞培养基+4nmol/L活化素A条件下培养3d,随后在StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+25μg/L表皮生长因子培养8d,中间每2的更换新鲜培养液1次;随后StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+10μg/L碱性成纤维细胞生长因子+0.5%胰岛素/转铁蛋白/硒诱导培养14d,间隔三天更换新鲜培养液。
多肽联合细胞因子诱导组为将干细胞先在StemCellMesenCultTM间充质干细胞培养基+4nmol/L活化素A+100μg/mL的多肽条件下培养3d,随后在StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+25μg/L表皮生长因子+100μg/mL的多肽培养8d,中间每2的更换新鲜培养液1次;随后StemCellMesenCultTM间充质干细胞培养基+10mmol/L尼克酰胺+10μg/L碱性成纤维细胞生长因子+0.5%胰岛素/转铁蛋白/硒+100μg/mL的多肽诱导培养14d,间隔三天更换新鲜培养液。
干细胞组:StemCellMesenCultTM间充质干细胞培养基培养干细胞等量时间。
取双硫腙粉末20mg溶于2mL二甲基亚砜中,0.22μm孔径滤膜过滤,取10μL加入1mL前述各组分化后的培养基中,37℃孵箱中孵育30min,PBS洗3次,显微镜下观察发现,细胞因子诱导组和多肽联合细胞因子组的细胞经双硫腙染色均可以形成红棕色胰岛样细胞团,而肽联合细胞因子组红棕色胰岛样细胞团密度更高,由于双硫腙可特异性结合胰岛素分子中的锌离子,这也表明,多肽能够更加有效的促进胰岛样细胞的分化。
诱导完成后,弃去培养液,PBS洗2次,加入L-DMEM(5.5mmol/L葡萄糖)孵育2h,收集培养液;诱导完成后,弃去培养液,PBS洗2次,加入H-DMEM(25mmol/L葡萄糖)孵育2h,收集培养液。严格按照人C肽定量ELISA试剂盒说明书,检测高糖(25mmol/L)刺激胰岛样细胞2h后C肽水平。结果如表1所示。
表1各组C肽水平
组别 | C肽(μg/L) |
细胞因子诱导组 | 0.71±0.12 |
多肽联合细胞因子诱导组 | 1.34±0.20 |
干细胞组 | 0 |
从表1可以看出,以低糖(5.5mmol/L)和高糖(25mmol/L)刺激胰岛样细胞,C肽水平在多肽联合细胞因子诱导组中的表达比细胞因子诱导组中的表达量要显著的提高,差异有显著性意义(P<0.05),这说明多肽能够促进Pdx-1的表达进而诱导胰岛样细胞的成熟及分泌C肽。
实施例5小鼠糖尿病治疗实验
将雄性C57BL/6J小鼠经适应性喂养1周后,随机取10只作为正常组,其余小鼠给予55mg/kg链脲佐菌素连续5d腹腔注射(过夜禁食12h后给药,给药后2h进食)。在给药后7,10d取尾静脉血检测血糖,2次都超过16.7mmol/L,即为1型糖尿病鼠。造模10d后,符合糖尿病标准小鼠随机分为3组:糖尿病组、尾静脉-多肽联合细胞因子诱导组诱导的胰岛样细胞组、尾静脉-细胞因子诱导组诱导的胰岛样细胞组,每组10只,每组尾静脉注射胰岛样细胞0.2mL胰岛样细胞悬液(含5×105个细胞)。正常组、糖尿病组注射生理盐水作为对照。移植后3周,第21d,各组小鼠禁食6h后,4%水合氯醛麻醉,眼球取血,检测血糖水平,同时离心血后取上清,严格按照人超敏胰岛素ELISA试剂盒说明书检测胰岛素水平。结果如表2所示。
表2各组血糖及胰岛素水平
从表2可以看出,正常组血糖水平远低于糖尿病组,差异有显著性意义
(P<0.01)。而二个治疗组尾静脉注射后的血糖与糖尿病组差异有显著性意义(P<0.05)。而且多肽联合细胞因子诱导组比细胞因子诱导组具有更好的降低血糖的效果。
正常组胰岛素水平和糖尿病组差异不显著。尾静脉注射后,与模型组相比,多肽联合细胞因子诱导组导致的分泌的胰岛素比细胞因子诱导组要高,差异有显著性意义(P<0.01)。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。最后应说明的是:以上各实施例仅用以说明本发明的技术方案,而非对其限制;尽管参照前述各实施例对本发明进行了详细的说明,但本领域的普通技术人员应当理解:其依然可以对前述各实施例所记载的技术方案进行修改,或者对其中部分或者全部技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (5)
1.一种培养脐带间充质干细胞的方法,其特征在于所述方法能够诱导脐带间充质干细胞分化为胰岛样细胞,包括:将脐带间充质干细胞先在StemCellMesenCultTM间充质干细胞培养基、4nmol/L活化素A、100μg/mL的多肽条件下培养3d,随后在StemCellMesenCultTM间充质干细胞培养基、10mmol/L尼克酰胺、25μg/L表皮生长因子、100μg/mL的多肽培养8d,中间每2的更换新鲜培养液1次;随后StemCellMesenCultTM间充质干细胞培养基、10mmol/L尼克酰胺、10μg/L碱性成纤维细胞生长因子、0.5%胰岛素/转铁蛋白/硒、100μg/mL的多肽诱导培养14d,间隔三天更换新鲜培养液,最后离心收集获得细胞即为诱导分化的胰岛样细胞,其中多肽为特异性诱导干细胞Pdx1高表达的多肽,其氨基酸序列如SEQ ID NO:1所示。
2.一种促进脐带间充质干细胞增殖及分化为胰岛样细胞的培养基,所述培养基包括如下各组:
培养基1:StemCellMesenCultTM间充质干细胞培养基、4nmol/L活化素A、100μg/mL多肽;
培养基2:StemCellMesenCultTM间充质干细胞培养基、10mmol/L尼克酰胺、25μg/L表皮生长因子、100μg/mL多肽;和
培养基3:StemCellMesenCultTM间充质干细胞培养基、10mmol/L尼克酰胺、10μg/L碱性成纤维细胞生长因子、0.5%胰岛素/转铁蛋白/硒、100μg/mL多肽;其中多肽为特异性诱导干细胞Pdx1高表达的多肽,其氨基酸序列如SEQ ID NO:1所示。
3.特异性诱导干细胞Pdx1高表达的多肽在促进脐带间充质干细胞增殖及分化的培养基中的应用,其中多肽的氨基酸序列如SEQ ID NO:1所示。
4.如权利要求3所述的用途,其中培养基中基础培养基为无血清干细胞培养基。
5.如权利要求3所述的用途,其中培养基中基础培养基为StemCell MesenCultTM间充质干细胞培养基。
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Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1637137A (zh) * | 2004-01-06 | 2005-07-13 | 中国人民解放军军事医学科学院野战输血研究所 | 一种诱导干细胞向胰岛样细胞分化的方法及其应用 |
CN105462913A (zh) * | 2014-09-05 | 2016-04-06 | 深圳市北科生物科技有限公司 | 一种诱导人脐带间充质干细胞分化为胰岛β细胞的方法 |
CN105624098A (zh) * | 2015-12-15 | 2016-06-01 | 深圳市第二人民医院 | PDX1蛋白诱导人脐带MSCs转分化为胰岛样细胞的方法 |
KR20160065325A (ko) * | 2014-11-28 | 2016-06-09 | 대한민국(농촌진흥청장) | Pdx1 유전자를 포함하는 벡터 및 이를 이용한 중간엽 줄기세포를 췌도 세포로 직접 분화시키는 방법 |
CN105670986A (zh) * | 2015-11-23 | 2016-06-15 | 王意忠 | 一种诱导人脐带间充质干细胞分化为胰岛样细胞的培养基及其诱导方法 |
CN106676056A (zh) * | 2016-11-08 | 2017-05-17 | 里程 | 诱导脐带间充质干细胞向胰岛素分泌样细胞分化的方法 |
CN107460158A (zh) * | 2017-08-10 | 2017-12-12 | 河南省银丰生物工程技术有限公司 | 一种诱导脐带间充质干细胞分化为形成胰岛素的方法 |
US20190264179A1 (en) * | 2016-11-08 | 2019-08-29 | Cheng Li | Serum-free medium inducing differentiation of umbilical cord mesenchymal stem cell into insulin-secretion-like cell and preparation method and use thereof |
CN113215094A (zh) * | 2021-05-17 | 2021-08-06 | 山东大学齐鲁医院 | 逆转2型糖尿病胰岛β细胞去分化的间充质干细胞外泌体及其制备方法和应用 |
CN114591910A (zh) * | 2022-03-28 | 2022-06-07 | 重庆市铂而斐细胞生物技术有限公司 | 一种利用低氧三维环境培养的间充质干细胞富集脐血造血干细胞的方法 |
-
2023
- 2023-07-04 CN CN202310810797.0A patent/CN116751735B/zh active Active
Patent Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1637137A (zh) * | 2004-01-06 | 2005-07-13 | 中国人民解放军军事医学科学院野战输血研究所 | 一种诱导干细胞向胰岛样细胞分化的方法及其应用 |
CN105462913A (zh) * | 2014-09-05 | 2016-04-06 | 深圳市北科生物科技有限公司 | 一种诱导人脐带间充质干细胞分化为胰岛β细胞的方法 |
KR20160065325A (ko) * | 2014-11-28 | 2016-06-09 | 대한민국(농촌진흥청장) | Pdx1 유전자를 포함하는 벡터 및 이를 이용한 중간엽 줄기세포를 췌도 세포로 직접 분화시키는 방법 |
CN105670986A (zh) * | 2015-11-23 | 2016-06-15 | 王意忠 | 一种诱导人脐带间充质干细胞分化为胰岛样细胞的培养基及其诱导方法 |
CN105624098A (zh) * | 2015-12-15 | 2016-06-01 | 深圳市第二人民医院 | PDX1蛋白诱导人脐带MSCs转分化为胰岛样细胞的方法 |
CN106676056A (zh) * | 2016-11-08 | 2017-05-17 | 里程 | 诱导脐带间充质干细胞向胰岛素分泌样细胞分化的方法 |
US20190264179A1 (en) * | 2016-11-08 | 2019-08-29 | Cheng Li | Serum-free medium inducing differentiation of umbilical cord mesenchymal stem cell into insulin-secretion-like cell and preparation method and use thereof |
CN107460158A (zh) * | 2017-08-10 | 2017-12-12 | 河南省银丰生物工程技术有限公司 | 一种诱导脐带间充质干细胞分化为形成胰岛素的方法 |
CN113215094A (zh) * | 2021-05-17 | 2021-08-06 | 山东大学齐鲁医院 | 逆转2型糖尿病胰岛β细胞去分化的间充质干细胞外泌体及其制备方法和应用 |
CN114591910A (zh) * | 2022-03-28 | 2022-06-07 | 重庆市铂而斐细胞生物技术有限公司 | 一种利用低氧三维环境培养的间充质干细胞富集脐血造血干细胞的方法 |
Non-Patent Citations (2)
Title |
---|
HUIJUAN YUAN等: "Regulation of mesenchymal stem cell differentiation and insulin secretion by differential expression of Pdx-1", MOL BIOL REP, 31 December 2012 (2012-12-31), pages 7777 - 7783 * |
王娟 等: "Pdx1基因修饰的人脐带间充质干细胞体外 诱导分化为胰岛β样细胞", 生物医学工程学杂志, vol. 28, no. 6, 31 December 2011 (2011-12-31), pages 1175 - 1180 * |
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