Umbilical cord and placental blood hematopoietic stem cells: collection, cryopreservation, and storage.

Human umbilical cord and placental blood is a rich source of hematopoietic stem cells that can be used to reconstitute hematopoiesis after intensive myeloablative therapy. Therefore, it has become necessary to develop techniques for (i) collecting large volumes of umbilical cord aseptically, (ii) optimizing the yield of viable cells after cryopreservation and thawing, and (iii) minimizing the risk of maternal T-cell contamination. Our protocol for the collection, cryopreservation, storage, and transport of human umbilical cord and placental blood is described in detail.     

猪苓多糖对脐血造血干细胞体外扩增及干细胞移植后免疫重建的调节效应

目的:人脐血造血干细胞得以移植于小鼠体内,其理论基础是放射线照射使小鼠骨髓枯竭,龛位腾出,为移植的脐血造血干细胞提供空间,而猪苓多糖是从中药猪苓中提取的多糖成分,具有免疫调节抗肿瘤作用,也是一种非T细胞促有丝分裂素,对放射线照射有一定的防护作用.观察猪苓多糖埘脐血造血干细胞体外扩增及干细胞移植后免疫重建效应.方法: 实验于2004-06/2006-05在兰州大学基础医学院免疫研究所完成.①实验材料:清洁级BALB/c小鼠由兰州生物制品研究所提供,8周龄,体质耸约20g,雌雄各半,实验过程中对动物处置符合动物伦理学标准.脐血样本由兰泰医院妇产科提供,产妇及家属对实验知情同意,并经医院伦理委员会批准.猪苓多糖由兰州大学第-医院提供.②实验方法:应用20,17.5,12.5,7.5 mg/L猪苓多糖体外扩增培养脐血造血干细胞,并测定细胞增殖率及CD34 细胞数.将BALB/c小鼠分为正常对照组:尾静脉、腹腔注射等量生理盐水:照射空白组:3.5C,y经60Coγ放射线照射联合尾静脉、腹腔注射等量生理盐水;照射药物组:3.5Gy经60Coγ放射线照射联合尾静脉注射等量生理盐水、腹腔庄射猪苓多糖;照射移植组(n=15):3.5Gy经60Coγ放射线照射联合脐血造血干细胞移植、腹腔注射等量生理盐水;照射移植药物组:35Gy经60Coγ放射线照射联合脐血造血干细胞移植、腹腔注射猪苓多糖.③实验评估:观察小鼠生存状况及血常规变化,流式细胞仪测定CD3、CD4、CD8、CD19水平.结果:57只小鼠均进入结果分析.①12.5 mg/L猪苓多糖增殖效率最为显著(P＜0.01),流式细胞仪测定显示CD34 细胞数扩增了6.33倍.移植药物组小鼠死亡率最低,死亡高峰集中在照射移植后7～14d左右.②移植药物组小鼠血象恢复正常,并且时间明显短于其他实验组(P＜0.05).③细胞及体液免疫恢复情况,移植药物组小鼠各项指标水平与正常小鼠无差别(P＞0.05),与其他各组小鼠比较,除CD4、CD19水平与照射移植小鼠无差别外均有差别(P＜0.05).结论:猪苓多糖对脐血造血干细胞有明显扩增作用,并能促进脐血造血干细胞移植小鼠免疫造血重建.     

Factors predicting the hematopoietic stem cells content of the umbilical cord blood

Umbilical cord blood (UCB) has been demonstrated to be alternative source of hematopoietic stem cells (HSCs). Unfortunately, the wide use of UCB Transplantation is limited due to the low number of HSCs. The aim of this study was to determine factors that affect the number of HSCs collected from UCB. 200 eligible donors were included for HSCs testing, including total nucleated cells (TNCs) and CD34+ cell number, by using univariate and multivariate analysis. In univariate analysis, factors positively associated with higher number of TNCs were maternal weight (P = 0.002), preeclampsia (P = 0.03), neonatal weight (P < 0.001), neonatal platelet count (P = 0.02), neonatal Rh (P = 0.03), gestational age (P = 0.04) and delivery type (P < 0.001). Factors positively associated with higher number of CD34+ cells were maternal weight (P < 0.007), preeclampsia (P = 0.02), maternal hypertension (P = 0.02) neonatal weight (P < 0.001), neonatal Rh type (P = 0.02) and delivery type (P = 0.04). In multivariate analysis, factors significantly influence TNCs were neonatal weight (P < 0.001), preeclampsia (P = 0.008), neonatal Rh type (P = 0.02) and delivery type (P < 0.001). While factors significantly influence number of CD34+ cells were maternal weight (P = 0.025), neonatal weight (P = 0.005), neonatal Rh (P = 0.006), nuchal cord (P = 0.026) and delivery type (P = 0.009). Conclusions factors significantly influence TNCs content of UCB were neonatal weight, preeclampsia, neonatal Rh and delivery type. While factors significantly influence number of CD34+ cells were maternal weight, neonatal weight, neonatal Rh, nuchal cord and delivery type.     

Issues in the quality of umbilical cord blood stem cells for transplantation

BACKGROUND: Because the frequency of umbilical cord blood (UCB) stem cell transplantation has increased, the quality of UCB available in banks is an important part of the success of UCB stem cell transplants. STUDY DESIGN AND METHODS: A quality assurance monitoring system was used to evaluate 268 UCB units provided to us for transplant by UCB banks in the United States and Europe. RESULTS: Quality issues were found in 151 (56%) of 268 units, and there were a total of 246 specific issues in 151 units. The issues involved quality control (54%), medical history (40%), and labels and documentation (6%). Risks to patients from these issues were likely in 10 percent, potential in 35 percent, and unlikely in 55 percent. CONCLUSION: Because standards have evolved over time, cord blood banks contain units that have different levels of quality. Some units have been placed in the usable inventory with incomplete test results and/or documentation or that may not meet the bank's own current criteria. Information about any quality or operating procedure deviation should be provided in sufficient detail and at the initiation of the search process so that transplant physicians can consider these quality issues against the unique value of a particular UCB unit.     

脐血造血干细胞移植中脐血输注的护理

脐血造血干细胞移植中如何使脐血造血干细胞顺利输注人病人体内是移植成功的重要环节,因此,脐血输注的护理尤为重要。笔阐述9例脐血造血干细胞移植患,脐血输注前输注时的护理,脐血输注后并发症的观察及护理3个部分的护理经验。9例患均成功进行了脐血输注。     

脐血间充质干细胞移植脊髓损伤恢复期患者的免疫功能变化

背景：间充质干细胞移植因其具有一定的免疫原性,对脊髓损伤患者免疫功能的影响尚无深入系统的报道。目的：观察脐血源间充质干细胞移植对脊髓损伤患者免疫功能的影响。方法：61例脊髓损伤患者采用静脉滴注结合腰椎脊髓蛛网膜下腔注射脐血间充质干细胞悬液治疗。应用流式细胞术及免疫比浊法分别测定患者移植前后T淋巴细胞亚群、免疫球蛋白及补体的含量。结果与结论：与治疗前相比,患者CD3＋、IgA及IgG有下降趋势,差异有显著性意义;CD4＋、CD8＋、CD4＋/CD8＋、IgM、C3及C4均有升高或下降趋势,但差异无显著性意义。提示脐血源间充质干细胞移植对细胞及体液免疫不会激活急性免疫应答,移植是安全的,存在负向免疫调节作用的可能,但尚需进一步完善相关指标,扩大样本,明确相关机制。     

脐血间充质干细胞移植脊髓损伤恢复期患者的免疫功能变化

背景:间充质干细胞移植因其具有一定的免疫原性,对脊髓损伤患者免疫功能的影响尚无深入系统的报道。目的:观察脐血源间充质干细胞移植对脊髓损伤患者免疫功能的影响。方法:61例脊髓损伤患者采用静脉滴注结合腰椎脊髓蛛网膜下腔注射脐血间充质干细胞悬液治疗。应用流式细胞术及免疫比浊法分别测定患者移植前后T淋巴细胞亚群、免疫球蛋白及补体的含量。结果与结论:与治疗前相比,患者CD3+、IgA及IgG有下降趋势,差异有显著性意义;CD4+、CD8+、CD4+/CD8+、IgM、C3及C4均有升高或下降趋势,但差异无显著性意义。提示脐血源间充质干细胞移植对细胞及体液免疫不会激活急性免疫应答,移植是安全的,存在负向免疫调节作用的可能,但尚需进一步完善相关指标,扩大样本,明确相关机制。     

Peripheral blood stem cell collection for allogeneic hematopoietic stem cell transplantation: Practical implications after 200 consequent transplants

Background

Proper stem cell mobilization is one of the most important steps in hematopoietic stem cell transplantation (HSCT). The aim of this paper is to share our 6 years’ experience and provide practical clinical approaches particularly for stem cell mobilization and collection within the series of more than 200 successive allogeneic HSCT at our transplant center.

脐血干细胞移植治疗脑卒中的研究现状

学术背景：目前传统的方法无法从根本上改善脑卒中后所造成的神经功能缺损，欲达到理想的恢复还赖于脑功能的重建。近年研究发现，脐血干细胞在特定诱导条件下可分化为神经细胞，修复受损的神经。 目的：总结脐血干细胞移植治疗脑卒中的研究进展。检索策略：由第一作者应用计算机检索PubMed数据库与万方数据库1995／2007年期间相关文献，检索词为“cord blood stem cells，Stroke，脐血干细胞，脑卒中”，并手工查阅相关书籍。对资料进行初审，选择脐血干细胞研究及采用脐血干细胞移植治疗脑卒中的研究，排除综述及重复文献。共检索到50篇相关文献。 文献评价：选择其中的31篇，文献的来源主要是脐血干细胞研究及采用脐血干细胞移植治疗脑卒中的动物及基础实验研究。 资料综合：脐血干细胞具有自我更新、增殖和多向分化潜能，在合适诱导下可分化为神经细胞。研究表明，脐血干细胞在体外诱导及体内移植能生成神经样细胞，能表达神经元表达神经元特异性标志物nestin、NF-M、NeuN、MAP2等。脐血干细胞治疗脑卒中在动物及实验研究基础上已取得了较大成就，但仍存在许多问题，细胞体外培养中分化增值的调控机制不十分清楚，临床研究报道甚少，诱导生成的神经细胞存活的时间,能否跟正常神经细胞一样发挥功能等，待进一步探讨。 结论：用来源丰富、免疫排斥概率小的脐血干细胞治疗多发的脑卒中，具有明显的优越性，为脑血管疾病患者神经功能的恢复带来新的希望。     

间充质干细胞在造血干细胞移植中的应用

应用计算机检索PubMed数据库2000—01／2008-01期间有关间充质干细胞与造血干细胞移植相关的文章，检索词为“MSC，HSCT”，限定文章语言种类为English。同时计算机检索维普数据库2000-01／2008-01期间相关文章，检索词为“间充质干细胞，造血干细胞移植”，限定文章语言种类为中文。并查阅与干细胞实验研究有关的书籍。对资料进行初审，选取符合要求的有关文章查找全文。纳入标准：①有关间充质干细胞对造血干细胞移植影响的研究。②造血干细胞移植机制的相关性研究。排除标准：重复性文章。共收集到66篇相关文献，28篇文献符合纳入标准。临床资料显示，两者共移植安全可行，可提高移植效果，降低近期死亡率。但其确切效果还需前瞻性随机对照临床实验进一步证实。     

Peripheral blood hematopoietic stem cell mobilization and collection efficacy is not an independent prognostic factor for autologous stem cell transplantation

BACKGROUND: The successful mobilization and collection of hematopoietic stem cells are dependent on a number of clinical factors such as previous chemotherapy and disease stage. The aim of this retrospective study was to determine whether the effectiveness of mobilization and collection is an independent prognostic factor for autologous stem cell transplantation outcome. STUDY DESIGN AND METHODS: A total of 358 patients who received transplants from January 2003 to December 2004 (201 male and 157 female patients, ages from 2.7 to 77.3 years with median of 53 years of age) underwent autologous hematopoietic stem cell collection after mobilization with granulocyte-colony-stimulating factor (G-CSF) or G-CSF plus chemotherapy priming. This retrospective study included patients with diagnoses of acute myelogenous leukemia, non-Hodgkin's lymphoma, Hodgkin's disease, multiple myeloma, and solid tumors. All patients underwent stem cell collection until a target or a minimum CD34+ cell dose was reached. Correlations were performed between stem cell mobilization and/or collection efficacy and transplantation outcomes. RESULTS: In general, both larger reinfused CD34+ cell dose and shorter number of days for the stem cell count to reach the minimum of 2 x 10(6) per kg CD34+ cells do not foster quicker engraftment. Reinfused CD34+ cell dose of less than 12 x 10(6) and number of days stem cell collection to reach this minimum CD34+ cell dose did not independently affect the overall survival (OS) or disease-free survival (DFS). CONCLUSION: The effectiveness of hematopoietic stem cell mobilization and collection as defined as number of days to reach a CD34+ cell dose of 2 x 10(6) per kg should not be used independently to forecast posttransplantation prognosis, engraftment, DFS, and OS.     

脐血间充质干细胞在造血干细胞移植中的临床应用

目的研究联合间充质干细胞和造血干细胞移植后的临床效果。方法实验组为联合移植的病例7例,对照组为外周造血干细胞移植的病例8例,通过分别观察每名患者的血小板重建时间、白细胞重建时间、住层流病房时间以及移植前后症状的改善情况、感染情况、GVHD发生率以及1年存活率来进行统计分析,判断2组的移植效果。结果实验组与对照组病例血小板重建平均时间分别为8.714、19.500 d,P<0.01;粒系重建平均时间分别为12.143、19.500 d,P<0.05;住层流病房平均时间分别为23.857、36.750 d,P<0.01;感染发生率分别分别为28.57%、62.50%;GVHD发生率为0、37.50%;一年内死亡率分别为0、12.5%。结论联合脐血间充质干细胞移植对造血干细胞移植后的归巢、发育、分化、成熟以及移植后的造血重建都有促进的作用,有助于减少移植后患者的感染率、GVHD发生率及死亡率,是造血干细胞移植技术的一步大跨越。     

ABO血型不合的异基因造血干细胞采集及移植效果研究

本研究评价COBE Spectra血细胞分离机按干细胞采集程序采集HLA配型相合、ABO血型不合供者外周血造血干细胞的效能,观察未去除红细胞和(或)血浆进行异基因外周血干细胞移植的效果。应用COBE Spectra血细胞分离机的自动干细胞采集程序采集28例异基因供者外周血干细胞,并选用同期ABO血型相合15例作对照。检测采集物有核细胞(NC)数、单个核细胞(MNC)比例及CD34+细胞计数,观察造血功能重建情况和转变为供者血型所需要的时间。结果表明,ABO血型不合和相合组采集物中的NC、CD34+细胞数、MNC比例无统计学差异(p>0.05)。ABO血型不合组和相合组中性粒细胞和血小板恢复的时间无统计学差异(p>0.05)。14例ABO血型主要不合患者,红系造血明显延迟,ABO血型不合组28名患者于移植后35-193天血型成功转变为供者型,和ABO血型相合组相比均有统计学差异(p<0.01)。结论:ABO血型不合不是异基因造血干细胞移植的障碍,主要不合可能是红系造血明显延迟的主要原因。     

单倍体相合造血干细胞联合脐带血间充质干细胞移植治疗急性重型再生障碍性贫血的疗效观察

本研究探讨单倍体相合造血干细胞移植联合脐带血间充质干细胞治疗重型再生障碍性贫血(SAA)的方法和疗效。对5例SAA的患者进行了单倍体相合造血干细胞移植。移植物选择单倍体相合供者骨髓或外周造血干细胞加脐带血间充质干细胞。观察移植后临床造血重建时间及近期并发症。结果显示,所有SAA患者移植后均获得造血重建,白细胞计数大于2×109/L的平均时间是13.8天,血小板计数大于20×109/L的平均时间是17.8天,第30天行患者外周血STR-PCR检测显示为完全供者的基因型。除1例发生癫痫失去联系外,其余4例均无病存活至今,仍在继续随访中。总之,单倍体相合造血干细胞联合脐带血间充质干细胞移植是治疗急性SAA有效可行的方法,但还须大样本的研究。     

造血微环境决定脐血造血干/祖细胞的早期分裂行为

目的 在特定生长因子的培养体系中，观察模拟造血微环境及粘附因素对人类脐血造血干/祖细胞早期分裂行为影响。方法 （1）应用流式细胞仪（FACS）分选CD34^ CD38^-细胞；（2）细胞培养体系中应用干细胞支持性基质AFT024及单一粘附因素纤维结合素（Fn)。并观察其对细胞早期分裂行为的影响。结果 （1）在联合生长因子的条件下，人类脐血CD34^ CD38^-细胞早期分裂呈现固定比例的静止，慢分裂，快分裂以及不对称分裂状态；（2）未观察到单一粘附因子Fn对其早期分裂行为的影响；（3）干细胞支持性基质AFT024所模拟的体外造血微环境可促使脐血造血干/祖细胞广泛增殖并更多地进行不对称分裂。结论 （1）CD34^ CD38^-细胞群体具有异质性。由具有不同增殖行为的亚群组成，在体外培养早期存在不对称分裂；（2）体外模拟的骨髓微环境AFT024基质滋养层较细胞因子及单一粘附因素能够更好地支持造血干/祖细胞的增殖并保持其自我更新的特性。     

1例神经母细胞瘤患儿脐带血造血干细胞移植术后的护理

研究发现,脐带血中含有可以重建人体造血和免疫系统的造血干／祖细胞,可用于造血干细胞移植,治疗多种疾病。造血干细胞移植整个过程是给予患者大剂量化、放疗等预处理后,将保存在-196摄氏度的液氮中经37～40摄氏度的温水解冻后快速输入患者体内,可以有效杀灭肿瘤细胞并重建造血系统,它的临床应用为恶性肿瘤患者提供了先进有效的治疗方法。本院于2009年10月收治了1例神经母细胞瘤的患儿,经采用造血干细胞移植后,取得较好的效果,现报道如下。