脐带脐血干细胞

摘要： 组织工程研究中的干细胞:培养与移植栏目设置1研究与报告1.1 骨髓干细胞1.2 造血干细胞1.3 脂肪干细胞1.4 脐带脐血干细胞1.5 肿瘤干细胞1.6 胚胎干细胞1.7 牙髓及牙周膜干细胞1.8 诱导性多能性干细胞2 技术与方法2.1 干细胞培养与分化2.2 干细胞移植2.3 干细胞外泌体2.4 干细胞因子及调控因子2.5 干细胞转基因表达2.6 干细胞与中药2.7 干细胞相关大数据分析.

摘要： 背景:骨关节炎是一种严重危害人类健康的慢性骨关节疾病.细胞治疗骨关节炎近年来发展较快,并得到了广泛关注.脐带来源的间充质干细胞治疗骨关节炎既具有其他来源间充质干细胞扩增分化容易,具有抗炎和招募作用等优势,又具有细胞年轻化、获得量大、无伦理问题、取材过程无痛苦、高增殖特性、多能分化特性、低免疫源性、无致瘤性等特点,是目前临床细胞治疗最常用的种子细胞.目的:拟利用同种异体脐带来源间充质干细胞治疗人膝关节骨关节炎,获得脐带来源间充质干细胞治疗骨关节炎的有效性和安全性数据,以期为干细胞治疗骨关节炎提供必要的理论基础和临床依据.方法:试验在中国北京市,北京大学人民医院关节病诊疗研究中心完成.在获得受试者的知情同意以及伦理委员会审批后(该试验前两部分均已注册(https://register.clinicaltrials.gov/),编号分别为 NCT03357770及NCT03358654,第三部分需要根据前两部分安全性评价后再行进行),按照既定的纳入/排除标准招募受试者.临床试验共分为3个部分:第一部分纳入3组受试者,每组3人,分别接受高、中、低3种等级的细胞剂量治疗,3组受试者在注射后进行随访,评价剂量限制性毒性,确立最大耐受剂量.第二部分即单臂临床试验,继续招募9名受试者进行干预试验,注射剂量为第一步试验确定的最高剂量,受试者在注射后进行随访,评价治疗的安全性及有效性.第三部分采用随机对照研究的试验设计,按照随机化的原则将受试者随机分为2组,每组7人,分别给予干细胞治疗和传统的透明质酸治疗,在试验期间,保证评价者盲、受试者盲以及干预者盲.在术后既定时间点进行随访,评价干预措施的安全性和疗效数据.主要终点是参考 NCI-CTCAE 标准,出现不明原因的局部及全身症状或死亡终止试验.预期结果:参考既往文献报道,经过该干预措施治疗后随访1至2年,受试者膝关节疼痛程度降低,关节功能评分提高,MRI检查见软骨缺损面积减小.预计试验时间共3年6个月.%BACKGROUND: Osteoarthritis (OA) is a kind of chronic bone and joint disease which seriously endangers human health. Cell therapy for OA has aroused widespread concern and gotten rapid development in recent years. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) have the advantages of easy amplification and differentiation, anti-inflammation and recruiting function such as MSCs from other sources. Furthermore, UC-MSCs are young cells that have large quantity, no ethical problems, high proliferative potential and pluripotent differentiation. UC-MSCs have been the most commonly used seed cells in clinical cell therapy. OBJECTIVE: To evaluate the efficacy and safety of UC-MSCs in the treatment of human knee OA to provide theoretical and clinical basis for stem cell therapy of OA. METHODS: The trail will be completed in Arthritis Clinic & Research Center, Beijing, China. Participants will be recruited according to established inclusion/exclusion criteria after obtaining the informed consent and the approval of the Ethics Committee (the first and second parts of the trial have been registered (https://register.clinicaltrials.gov/), with the identifier No. NCT03357770 and NCT03358654, and the third part will be carried out according to the conclusion of the first and second parts). The clinical trial will be divided into three parts: in the first part three groups will be recruited. Each group will contain three participants. The three groups of participants will be treated with high, medium and low dose of MSCs, respectively. Participants will be followed up to evaluate dose-limiting toxicity so as to determine the maximum tolerated dose. The second part will be a single-arm clinical trial. Nine participants will be recruited. The injection dose will be the maximum tolerated dose determined in the first part. Participants will be followed up to evaluate the safety and efficacy of the treatment. The third part will be a randomized controlled trial. Participants will be randomly divided into two groups (n=7 per group) and treated with MSCs and hyaluronic acid, respectively. During the trial, evaluators, participants and interveners will be unaware of grouping information and interventions. Participants will be followed up at designed time points after treatment to evaluate the safety and efficacy of the intervention. The trial will be terminated if there are unexplained local and systemic symptoms or death according to the NCI-CTCAE criteria. EXPECTED RESULTS: With reference to the previous literature, the knee pain will be relieved, the score of knee joint function will increase, and the cartilage defect area will decrease on MRI at 1-2 years after the intervention. The trail is expected to spend 3 years and 6 months.

摘要： BACKGROUND: Studies have demonstrated that platelet-rich plasma can promote osteogenesis and proliferation of bone marrow mesenchymal stem cells, adipose-derived stem cells and skeletal muscle satellite cell, but whether it can promote the proliferation and osteogenesis of human umbilical cord mesenchymal stem cells (hUC-MSCs) is unclear. OBJECTIVE: To investigate the effects of different concentrations of platelet-rich plasma on the proliferation and osteogenic differentiation of hUC-MSCs. METHODS: Passage 5 hUC-MSCs were cultured in medium containing different concentrations of platelet-rich plasma (0, 250, 500, 750, 1000, 2000 ng/L), respectively. Cell proliferation was detected at 1, 3, 5, 7 days after culture, and the best platelet-rich plasma mass concentration was screened. Afterwards, passage 5 hUC-MSCs were divided and cultured in complete medium (blank control group), optimal concentration of platelet-rich plasma (platelet rich plasma group), osteogenesis induction medium (osteogenic induction group), or the osteogenesis induction medium containing the optimal concentration of platelet-rich plasma (combined group). Cell activity of alkaline phosphatase was detected after cultivated for 3, 7, 14 days. Osteopontin, bone specific transcription factor, and osteocalcin mRNA relative expression levels were detected after cultivated for 7, 14, 21 days. Mineralization of the extracellular matrix was detected after cultivated for 21 days. RESULTS AND CONCLUSION: After 5 and 7 days of culture, the cell proliferation was higher in 500, 750, 1000 ng/L platelet-rich plasma groups than 0 ng/L group (P < 0.05), and 750 ng/L platelet-rich plasma showed the best effect on cell proliferation, which was used in the following experiments. Compared with the other groups, the combined group had significantly increased alkaline phosphatase activity (P < 0.05), and up-regulated osteopontin, bone specific transcription factor and osteocalcin mRNA relative expression levels (P < 0.05) at different culture times. In addition, the degree of extracellular matrix mineralization in the combined group was also higher than that in the other three groups (P < 0.05).To conclude, 750 ng/L platelet-rich plasma can promote hUC-MSCs proliferation and osteogenic differentiation.%背景:研究证明,富血小板血浆可促进骨髓间充质干细胞、脂肪来源干细胞及骨骼肌卫星细胞的增殖及成骨分化,但其是否可促进人脐带间充质干细胞的增殖及成骨分化目前尚不清楚.目的:观察不同质量浓度富血小板血浆对体外培养人脐带间充质干细胞增殖及成骨分化的影响.方法:将第5代人脐带间充质干细胞分6组培养,分别以含0,250,500,750,1000,2000 ng/L活化富血小板血浆的培养基中培养,1,3,5,7 d后,检测细胞增殖,筛选最佳富血小板血浆质量浓度,进行以下实验.将第5代细胞人脐带间充质干细胞分4组培养,空白对照组添加完全培养基,富血小板血浆组添加最佳浓度的富血小板血浆,成骨诱导组添加成骨诱导培养基,联合组添加最佳浓度的富血小板血浆与成骨诱导培养基,培养3,7,14 d,检测细胞碱性磷酸酶活性;培养7,14,21 d,检测骨标志基因骨桥蛋白、骨特异性转录因子、骨钙蛋白mRNA的相对表达量;培养21 d,检测细胞外基质的矿化.结果与结论:①培养5,7 d时,500,750,1000 ng/L富血小板血浆组的细胞增殖高于0 ng/L富血小板血浆组(P<0.05),其中以750 ng/L质量浓度促增殖效果最明显,以此浓度进行以下实验;②联合组培养不同时间点的碱性磷酸酶活性高于其余3组(P<0.05);③联合组培养不同时间点的骨桥蛋白、骨特异性转录因子、骨钙蛋白mRNA相对表达量高于其余3组(P<0.05);④联合组细胞外基质矿化程度高于其余3组(P<0.05);⑤结果表明,750 ng/L富血小板血浆可促进人脐带间充质干细胞的增殖及成骨分化.

摘要： 背景:脐带间充质干细胞治疗肝衰竭已经取得了一定的疗效,但是肝衰竭患者的血清微环境对脐带间充质干细胞分化的影响尚未见报道.目的:探讨慢加急性肝衰竭患者血浆置换前、后血清对脐带间充质干细胞向功能肝细胞分化的影响.方法:采用组织块贴壁培养法获得脐带间充质干细胞并进行细胞形态及表型鉴定;取第3代脐带间充质干细胞,用含有肝衰竭患者血浆置换前、后血清的 α-MEM培养基诱导培养,常规胎牛血清培养基为对照组,显微镜下观察各组细胞的形态,免疫组化检测细胞甲胎蛋白、白蛋白和细胞角蛋白18的表达水平.结果与结论:①组织块贴壁培养法能够获得大量高纯度的脐带间充质干细胞,细胞高表达CD44,CD73,CD90,CD105,不表达CD45,符合间充质干细胞的表型特征;②慢加急性肝衰竭患者血清可改变脐带间充质干细胞的形态,并诱导脐带间充质干细胞表达甲胎蛋白、白蛋白和细胞角蛋白18.血浆置换后血清组3种蛋白阳性率高于血浆置换前血清组,差异有显著性意义(P<0.05);③结果表明,肝衰竭患者血浆置换后血清更加有利于脐带间充质干细胞生长及向肝细胞分化.%BACKGROUND: Umbilical cord mesenchymal stem cells (UC-MSCs) have made certain curative effect on hepatic failure, but little is reported on the effect of hepatic failure patient's serum microenvironment on UC-MSCs differentiation ability. OBJECTIVE: To investigate the effect of hepatic failure patient's serum on the differentiation of umbilical cord mesenchymal stem cells into hepatic cells. METHODS: UC-MSCs were isolated by tissue adherent method and the cell morphology and phenotype identified by microscope and flow cytometry. Alpha-MEM media with serum before/after plasmapheresis were used to culture the hirdgeneration of UC-MSCs, and regular fetal bovine serum culture medium acted as control group. Inverted microscope was used to observe the cell morphology in three groups. Immunohistochemical method was used to measure expression level of alpha fetoprotein, albumin and cytokeratin 18. RESULTS AND CONCLUSION: A great amount of high-purity UC-MSCs could be obtained using tissue adherent method, which highly expressed CD44, CD73, CD90, CD10, but did not express CD45. Hepatic failure patient's serum could change the morphology of UC-MSCs and induce UC-MSCs to express alpha fetoprotein, albumin and cytokeratin 18. The positive expression of alpha fetoprotein, albumin and cytokeratin 18 was significantly increased after plasmapheresis (P < 0.05). To conclude, hepatic failure patient's serum after plasmapheresis exert more benefits to induce UC-MSCs to differentiate into hepatic cells than that before plasmapheresis.

摘要： 背景:近年来间充质干细胞的免疫调节作用可被应用于诱导机体免疫耐受研究.目的:评估脐带间充质干细胞应用于肝移植患者的安全性和可行性.方法:培养人脐带间充质干细胞并进行鉴定.经医学伦理委员会同意后,将总计50例患者经1:1的比例按入组时间随机分入试验组和对照组,每组25例.试验组分别在术中门静脉灌注及术后第3天给予颈内静脉输注脐带间充质干细胞,剂量为1×106/kg(制备成50 mL的细胞悬液).两组患者均采用标准免疫抑制方案.术前,术后第3,7天,术后第1,2,3,6,12个月检测血生化、免疫功能指标,统计急、慢性排斥反应发生率,感染发生率,移植相关并发症发生率.结果与结论:①与对照组相比,术后第3,7天试验组外周血的CD4+CD25+(调节性T细胞)细胞比例高于对照组,差异有显著性意义(P0.05);③试验组术后肝功能异常事件的发生率明显低于对照组,差异有显著性意义(P0.05);⑤以上结果提示,肝移植术后静脉输注脐带间充质干细胞安全可行,早期可促进CD4+CD25+(调节性T细胞)的增殖和活化,降低CD4+(辅助性/诱导性T细胞)比例及CD4+/CD8+T淋巴细胞的比值,进而改善肝移植受者免疫状态.%BACKGROUND: In recent years, the immunoregulatory effects of mesenchymal stem cells (MSCs) can be used to induce immune tolerance. OBJECTIVE: To evaluate the safety and feasibility of human umbilical cord-derived MSCs (hUC-MSCs) in liver transplantation patients. METHODS: hUC-MSCs were cultured and identified. After approved by the Medical Ethics Committee, a total of 50 patients were randomly divided into experimental group and control group according to the proportion of 1:1. In the experimental group, hUC-MSCs were perfused by the portal vein during the operation and infused into the jugular vein on the 3rd day after the operation. The injection dose was 1×106/kg (prepared as 50 mL of cell suspension). Both groups received standard immunosuppressive regimens. Blood biochemistry and immune function indicators were detected preoperatively and at postoperative days 3, 7, months 1, 2, 3, 6, 12. Acute and chronic rejection rates, incidence of infection, and incidence of transplantation-related complications were recorded. RESULTS AND CONCLUSION: (1) At 3 and 7 days after the operation, the percentage of peripheral blood CD4+CD25+ cells (regulatory T cells) in the experimental group was significantly higher than that in the control group (P 0.05). (3) The incidence of abnormal liver function in the experimental group was significantly lower than that in the control group (P 0.05). Overall, the intravenous infusion of hUC-MSCs is safe and feasible in liver transplantation patients, which in early stage can promote the the proliferation and activation of CD4+CD25+ cells (regulatory T cells), reduce the percentage of CD4+ cells (helper/inducer T cells) and lower the ratio of CD4+/CD8+ T cells, thereby improving the immune status in liver transplantation patients.

摘要： 背景:根据体外培养的成体干细胞自发恶性转化现象及肿瘤干细胞理论,人脐带间充质干细胞在体外培养过程中可能产生变异,体内移植后可能存在致瘤性风险.因此,建立健全的体外安全性检测程序,将在干细胞临床应用上起积极推动作用.目的:探讨人脐带间充质干细胞致瘤性机制及DNA甲基化转移酶的表达水平.方法:采用组织块贴壁法分离、培养、扩增出人脐带间充质干细胞,利用3-甲基胆蒽促使人脐带间充质干细胞发生恶性转化,通过形态学观察,裸鼠成瘤实验进行验证,成瘤组织进行病理学鉴定、原代细胞培养,荧光定量PCR检测人脐带间充质干细胞与肿瘤细胞DNA甲基化转移酶表达水平的差异.结果与结论:①人脐带间充质干细胞经3-甲基胆蒽处理后形成恶性转化细胞,呈恶性生长,细胞核型呈非整数倍体改变,注入裸鼠体内后能形成恶性肿物;②经荧光定量PCR检测,恶性转化后肿瘤细胞(实验组)相比二甲基亚砜培养人脐带间充质干细胞(对照组),其DNA甲基化转移酶的基因表达量明显增高;③结果表明,人脐带间充质干细胞在一定条件下能发生恶性转化,包括形态学变化及表观遗传学层面改变,DNA甲基化转移酶可作为预防干细胞移植后成瘤的进一步研究方向.%BACKGROUND:Human umbilical cord mesenchymal stem cells (hUC-MSCs) may be mutated duringin vitro culture based on the spontaneous malignant transformation of adult stem cells and tumor stem cell theory, and there may be a risk of tumorigenesis after in vivo transplantation. Therefore, to establish and perfect the in vitro safety testing procedures will actively promote the clinical application of stem cells. OBJECTIVE:To investigate the tumorigenic mechanism of hUC-MSCs and the expression level of DNA methyltransferase (DNMTs) in hUC-MSCs. METHODS:Primary hUC-MSCs were isolated and expanded by tissue adherent culture. 3-Methycholanthrene was used to cause the malignant transformation in hUC-MSCs (experimental group), followed by morphological observation and tumorigenesis experiment in nude mice. Then, the tumor tissues were obtained and identified by pathological examination and primary cell culture, and the levels of DNMTs mRNA in hUC-MSCs treated with 3-methycholanthrene and dimethyl sulfoxide (control group) were detected by real-time RT-PCR and compared. RESULTS AND CONCLUSION:hUC-MSCs treated with 3-methycholanthrene led to malignant transformation, which showed malignant growth and non-integer ploidy changes in the cell nuclei, and formed a malignant tumor in immune-deficient mice after injection. Compared with the control group, the cells in the experimental group showed higher expression of DNMTs mRNA as detected by real-time RT-PCR. To conclude, hUC-MSCs can trigger malignant transformation in the morphology and the epigenetics under certain conditions. DNMTs can be a candidate for prevention against malignant transformation of transplanted stem cells.

摘要： 背景:人脐带华通胶间充质干细胞移植到犬退变椎间盘模型中可存活,恢复部分椎间盘高度,但其对炎性损伤后髓核细胞是否有修复治疗作用目前还没有文献报道.目的:观察人脐带华通胶间充质干细胞对白细胞介素1β诱导大鼠髓核细胞退变的保护作用.方法:将第3代SD大鼠髓核细胞分3组培养,对照组用含胎牛血清的培养基培养24 h,然后换完全培养基培养24 h;白细胞介素1β组以含白细胞介素1β和胎牛血清的培养基培养24 h,然后换完全培养基培养24 h;实验组先以含白细胞介素1β和胎牛血清的培养基培养24 h,再更换完全培养基与人脐带华通胶间充质干细胞非直接共培养24 h.采用Annexin V/PI染色后流式细胞仪测定3组髓核细胞凋亡率,RT-PCR测定各组髓核细胞基质金属蛋白酶4、基质金属蛋白酶3、基质金属蛋白酶抑制剂1的表达,caspase试剂盒检测caspase-3、caspase-8和caspase-9活性.结果与结论:①白细胞介素1β组、实验组细胞凋亡率高于对照组(P co-culture group > normal control group, and there were significant differences between groups (P < 0.05). Compared with the control group,the expression levels of ADAMTS-4 and MMPs-3 were significantly higher and the expression level of TIMP-1 was significantly lower in the intervention and co-culture groups (P < 0.05). Compared with the intervention group, the expression levels of ADAMTS-4 and MMPs-3 were significantly lower and the expression level of TIMP-1 was significantly higher in the co-culture group (P < 0.05). The expression activities of caspase-3, caspase-8 and caspase-9 were highest in the intervention group, followed by the co-culture group, and lowest in the normal control group, and there were significant differences between groups (P < 0.05). To conclude, hWJCs has obviously inhibitory effect on the apoptosis of degenerated nucleus pulposus cells induced by interleukin-1β.

摘要： 背景：间充质干细胞是体内微环境的重要组成部分且影响肿瘤细胞的多种生物学行为，间充质干细胞的潜在临床价值是近年来的研究热点。目的：通过基因芯片方法分析脐带间充质干细胞对急性早幼粒细胞白血病细胞系NB4细胞基因组表达谱的影响。方法：体外建立脐带间充质干细胞与急性早幼粒细胞白血病细胞系NB4细胞共培养体系，检测脐带间充质干细胞对NB4细胞增殖、凋亡和分化的影响，分别提取了NB4细胞单独培养组和NB4细胞+脐带间充质干细胞共培养组NB4细胞的mRNA并将其反转录为cDNA，两组分别标记荧光，做成探针并与基因芯片杂交，检测其荧光强度，分析两组基因表达谱的差异。结果与结论：①脐带间充质干细胞促进NB4细胞的增殖和分化并抑制其凋亡；②脐带间充质干细胞作用后的NB4细胞的基因表达谱中有530个基因上调和53个基因下调，与基因相关的功能和信号通路也在一定程度上受到了调节；③结果表明，脐带间充质干细胞通过改变肿瘤细胞内大量基因表达、基因功能及多种信号通路影响NB4细胞的生物学行为。

摘要： 背景：人脐带华通胶间充质干细胞移植到犬退变椎间盘模型中可存活，恢复部分椎间盘高度，但其对炎性损伤后髓核细胞是否有修复治疗作用目前还没有文献报道。目的：观察人脐带华通胶间充质干细胞对白细胞介素1β诱导大鼠髓核细胞退变的保护作用。方法：将第3代SD大鼠髓核细胞分3组培养，对照组用含胎牛血清的培养基培养24h，然后换完全培养基培养24h；白细胞介素1β组以含白细胞介素1β和胎牛血清的培养基培养24h，然后换完全培养基培养24h；实验组先以含白细胞介素1β和胎牛血清的培养基培养24h，再更换完全培养基与人脐带华通胶间充质干细胞非直接共培养24h。采用AnnexinV/PI染色后流式细胞仪测定3组髓核细胞凋亡率，RT-PCR测定各组髓核细胞基质金属蛋白酶4、基质金属蛋白酶3、基质金属蛋白酶抑制剂1的表达，caspase试剂盒检测caspase-3、caspase-8和caspase-9活性。结果与结论：①白细胞介素1β组、实验组细胞凋亡率高于对照组(P<0.05)，实验组低于白细胞介素1β组(P<0.05)；②白细胞介素1β组、实验组基质金属蛋白酶4、基质金属蛋白酶3表达量高于对照组(P<0.05)，基质金属蛋白酶抑制剂1表达量低于对照组(P<0.05)；实验组基质金属蛋白酶4、基质金属蛋白酶3表达量低于白细胞介素1β组，基质金属蛋白酶抑制剂1表达量高于白细胞介素1β组(P<0.05)；③白细胞介素1β组、实验组caspase-3、caspase-8、caspase-9相对活性高于对照组(P<0.05)，实验组caspase-3、caspase-8、caspase-9相对活性低于白细胞介素1β组(P<0.05)；④结果表明，人脐带华通胶间充质干细胞对白细胞介素1β诱导的髓核细胞退变凋亡有明显的抑制作用。

摘要： BACKGROUND: Mesenchymal stem cells (MSCs) are an important component of the in vivo microenvironment and act on multiple biological behaviors of tumor cells. The potential clinical value of MSCs has become an issue of concern in recent years.OBJECTIVE: To investigate the gene expression profiles of acute promyelocytic leukemia (APL) cell line NB4 treated with umbilical cord-derived MSCs (UC-MSCs) using cDNA microarray.METHODS: In vitro co-culture system was constructed, and then cellular proliferation, apoptosis and differentiation status of NB4 cells treated with UC-MSCs were evaluated. Two cDNA probes were prepared through reverse transcription from mRNA of NB4 cells treated with or without UC-MSCs. The probes were labeled with fluorescence dyes individually, hybridized with cDNA microarray, and their fluorescent intensities were scanned. The genes were screened through the analysis of the difference in two gene expression profiles.RESULTS AND CONCLUSION: UC-MSCs promoted the proliferation and differentiation, while reduced the apoptosis of NB4 cells. The analysis of gene expression profiles indicated that after co-culture with UC-MSCs, 530 genes were up-regulated and 53 genes were down-regulated. Accordingly, specific gene function and pathway signaling related were also regulated to some extent. Overall, UC-MSCs influence can major biological behaviors of NB4 cells by changing expression of a large amount of genes, gene-related function and multiple intracellular signaling pathways.%背景:间充质干细胞是体内微环境的重要组成部分且影响肿瘤细胞的多种生物学行为,间充质干细胞的潜在临床价值是近年来的研究热点.目的:通过基因芯片方法分析脐带间充质干细胞对急性早幼粒细胞白血病细胞系NB4细胞基因组表达谱的影响.方法:体外建立脐带间充质干细胞与急性早幼粒细胞白血病细胞系NB4细胞共培养体系,检测脐带间充质干细胞对NB4细胞增殖、凋亡和分化的影响,分别提取了NB4细胞单独培养组和NB4细胞+脐带间充质干细胞共培养组NB4细胞的mRNA并将其反转录为cDNA,两组分别标记荧光,做成探针并与基因芯片杂交,检测其荧光强度,分析两组基因表达谱的差异.结果与结论:①脐带间充质干细胞促进NB4细胞的增殖和分化并抑制其凋亡;②脐带间充质干细胞作用后的NB4细胞的基因表达谱中有530个基因上调和53个基因下调,与基因相关的功能和信号通路也在一定程度上受到了调节;③结果表明,脐带间充质干细胞通过改变肿瘤细胞内大量基因表达、基因功能及多种信号通路影响NB4细胞的生物学行为.

摘要： 本发明涉及从脐带血中分离和培养间充质干细胞/祖细胞的方法，还涉及将脐带血来源的间充质干细胞/祖细胞分化成各种间充质组织的方法。所述方法包括如下步骤：将脐带血覆盖在Ficoll-Hypaque溶液上；将Ficoll-Hypaque溶液上的脐带血离心，获得单核细胞层；使通过单核细胞单层培养获得的细胞与针对间充质干细胞/祖细胞特异性抗原的抗体反应预定的孵育期；使用细胞分选仪仅分离与其相应抗体相结合的细胞；将分离的细胞培养，从而获得具有高纯度和极好生存能力的间充质干细胞/祖细胞。本发明的间充质干细胞/祖细胞能够分化成包括软骨细胞和成骨细胞在内的各种间充质组织。因此，根据本发明的细胞分离和培养方法能够实现间充质干细胞/祖细胞的大规模生产，本发明获得的细胞在受损间充质组织的再生和治疗上是有用的。

摘要： 本发明涉及细胞培养技术领域，尤其涉及一种以自身脐带间充质干细胞作为基质细胞扩增人类脐带血造血干细胞的方法。本发明提供了一种体外大量扩增CB‑HSCs的方法，即利用来自自身的脐带间充质干细胞(umbilical cord mesenchymalstem cells，UCMSC)作为基质细胞与CB‑HSCs共培养，从而模拟CB‑HSCs在体内的生长微环境，用此方法可有效扩增CB‑HSCs，而且扩增后的细胞克服了细胞因子扩增法的易于损伤、崩解等缺陷。

摘要： 本发明提供了包含脐带血衍生的间充质干细胞的组合物用于诱导神经前体细胞或神经干细胞分化增殖为神经细胞的用途，所述组合物对于治疗神经损伤性疾病是有效的。

摘要： 本发明涉及一种脂肪干细胞在促进脐带血造血干细胞增殖的方法。本发明通过筛选和优化获得能够特异性促进脐带血干细胞中FOXO3高表达的促进活性肽，该多肽能够通过提高FOXO3的表达，来促进脐带血干细胞的增殖以及向红细胞的分化；还提供了一种改进的采用脂肪干细胞来提高脐带血干细胞增殖的方法，所述方法包括将脂肪干细胞与脐带血干细胞一起进行培育而获得。将扩增的造血干细胞进一步的在特制的诱导培养基中培养能够获得分化的红细胞。本发明能够高速度的扩增造血干细胞并且能够获得红系特性较好的红细胞，可以大规模用于后续的临床研究，具有较好的应用价值。

摘要： 本发明提供了SphK2抑制剂ABC294640促进脐带血体外扩增和增加脐带血移植后造血重建和免疫重建速度的用途。ABC294640可以有效增加脐带血中造血干数量和功能可用于脐带血干细胞的体外培养增加造血干细胞的数量，也可用于脐带血干细胞骨髓移植后加快脐带血移植后造血重建的速度和效率。本发明可应用于脐带血的扩增、存储以及脐带血干细胞移植后的治疗。

摘要： 本发明涉及从脐带血中分离和培养间充质干细胞/祖细胞的方法，还涉及将脐带血来源的间充质干细胞/祖细胞分化成各种间充质组织的方法。所述方法包括如下步骤：将脐带血覆盖在Ficoll－Hypaque溶液上；将Ficoll－Hypaque溶液上的脐带血离心，获得单核细胞层；使通过单核细胞单层培养获得的细胞与针对间充质干细胞/祖细胞特异性抗原的抗体反应预定的孵育期；使用细胞分选仪仅分离与其相应抗体相结合的细胞；将分离的细胞培养，从而获得具有高纯度和极好生存能力的间充质干细胞/祖细胞。本发明的间充质干细胞/祖细胞能够分化成包括软骨细胞和成骨细胞在内的各种间充质组织。因此，根据本发明的细胞分离和培养方法能够实现间充质干细胞/祖细胞的大规模生产，本发明获得的细胞在受损间充质组织的再生和治疗上是有用的。

摘要： 本发明涉及细胞培养技术领域，尤其涉及一种以自身脐带间充质干细胞作为基质细胞扩增人类脐带血造血干细胞的方法。本发明提供了一种体外大量扩增CB‑HSCs的方法，即利用来自自身的脐带间充质干细胞(umbilical cord mesenchymalstem cells，UCMSC)作为基质细胞与CB‑HSCs共培养，从而模拟CB‑HSCs在体内的生长微环境，用此方法可有效扩增CB‑HSCs，而且扩增后的细胞克服了细胞因子扩增法的易于损伤、崩解等缺陷。

摘要： 本发明涉及干细胞制剂技术领域，特别涉及一种脐带间充质干细胞活性物与脐带血干细胞活性物的复合物的制备方法，提取脐带组织得到原代脐带干细胞；培育第五代原代脐带间充质干细胞，并获得细胞培养上清液进行离心，并对所述上清溶液进行过滤、超滤浓缩，加入冻干保护剂，并进行真空冷冻干燥，得到脐带间充质干细胞活性物冻干粉末；获得脐带血，进行传代培养；分离得到脐带血干细胞溶液，反复冻融至细胞完全破碎，得到脐带血干细胞提取物溶液，冻干制得脐带血干细胞活性物冻干粉。本发明所得的制剂具有更好促进皮肤烫伤修复的作用，能促进皮肤烫伤的创面愈合、更好地促进其表皮的再生化、皮肤细胞增殖再生能力等作用。

摘要： 本发明提供了包含脐带血衍生的间充质干细胞的组合物用于诱导神经前体细胞或神经干细胞分化增殖为神经细胞的用途，所述组合物对于治疗神经损伤性疾病是有效的。

摘要： 本发明公开了一种脐带血造血干细胞分离设备、干细胞培养选型方法及其过敏性鼻炎上的应用，属于造血干细胞领域，一种脐带血造血干细胞分离设备，通过内封取样管的设置，分离时，在取样之前，先挤压主袋，使血液浸满内封取样管下端部，可手动按压内封取样管顶部，使自吸液球内负压的状态被破坏，从而直接吸收内封取样管内血液，之后再次按压内封取样管顶部，使自吸液球与主袋之间再次密封隔离，此时，在通过注射器扎入自吸液球内取样，相较于现有技术，取样时，完全与主袋内血液隔离，进而有效避免内部血液被污染的情况发生，有效降低对干细胞后续保存或应用的影响。