脐带血移植的应用进展及脐带血库建设

脐带血(umbilical cord blood)作为公认的造血干细胞重要来源之一,已经被广泛地用于治疗儿童和成人的良恶性血液系统疾病以及中枢神经系统疾病、实体瘤、缺血性下肢血管病和组织再生等。相对于骨髓移植和外周血来源的造血干细胞移植,脐带血移植(UCBT)在细胞收集使用、干细胞增殖能力以及移植物抗宿主反应等方面都具有明显的优势。目前的数据显示,因为HLA配型等原因而无法进行骨髓移植的患者应该尽早进行UCBT。此外,UCBT的增多促进了脐带血库的快速建设。本文针对UCBT和脐带血库的最新进展进行了综述。     

New allergies after cord blood transplantation

Background aimsUmbilical cord blood transplantation (CBT) is an effective treatment for benign and malignant diseases. Late effects of CBT are not well described in the literature. In the present study, we present our experience of new-onset allergies in long-term survivors after CBT.MethodsAfter an initial patient had a severe peanut allergic reaction after CBT, all CBT patients were prospectively followed for new allergy development. Fifty patients received CBT between March 2006 and June 2011.ResultsThe median follow-up after CBT was 447 days (range, 12–2022). At the time of analysis, 30 patients were alive, with 3-year survival of 55.5%; median follow-up of surviving patients was 910 days (range, 68–2022). The allergic syndrome developed in five patients, with the cumulative incidence of new allergies at 2 years of 18.4% (95% confidence interval, 10.8–26). The median time to onset of new allergy after transplantation was 298 days (range, 250–809).ConclusionsAllergy development has been linked to a delayed maturation of the immune system in several studies. We present the first case series of patients who had new allergies after CBT. Further study of this novel complication as well as counseling of patients after CBT would be important.     

脐带血干细胞移植治疗非恶性血液病进展

异基因造血干细胞移植（allo-HSCT）是治愈多种非恶性病的有效方法。脐带血干细胞（UCB）具有免疫原性低、人类白细胞抗原不合耐受性好、移植物抗宿主反应发生率低以及获取相对快捷等特点,可作为非恶性血液疾病患者allo-HSCT的来源。本文简要综述脐血干细胞移植在原发性免疫缺陷病、遗传性骨髓衰竭、遗传代谢病以及自身免疫性疾病等非恶性血液疾病的治疗效果。     

脐血干细胞移植对儿童重组活化基因突变重症联合免疫缺陷病疗效分析

目的探讨脐血干细胞移植(UCBT)治疗儿童重组活化基因(RAG)突变重症联合免疫缺陷病(SCID)的疗效。方法回顾性分析2015年1月至2019年6月于复旦大学附属儿科医院明确诊断为RAG突变并接受UCBT的8例SCID患儿,其中男6例,女2例。所有患儿均接受白消安(BU)/环磷酰胺(CY)为主的减低毒性预处理方案,采用他克莫司单药预防移植物抗宿主病(GVHD),收集脐血干细胞移植前后的临床及实验室资料,评估患儿治疗效果。不符合正态分布的连续性变量资料用中位数(最小值~最大值)表示,分类资料用例数(相对比)表示。结果8例接受UCBT的RAG突变患儿分析结果显示,脐带血输注中位总有核细胞数为14.73(5.06~27.27)×10^7个/kg,中位CD34+细胞数为4.14(1.75~13.30)×10^5个/kg。8例患儿中7例成功植入,中性粒细胞植入时间25(15~45)d,血小板植入时间34(33~43)d。中位随访时间9.2(0.3~31.3)个月。3例患儿移植早期死亡,5例患儿无病生存并获得免疫重建,脱离静脉丙种球蛋白输注的时间为84(63~100)d。结论UCBT治疗RAG突变所致SCID取得一定疗效。     

Low rate of infusional toxicity after expanded cord blood transplantation

Background aimsUmbilical cord blood (CB) is used with increasing frequency to restore hematopoiesis in patients with bone marrow transplant who lack a suitable human leukocyte antigen–matched donor. CB transplantation is limited by low cell doses and delays in neutrophil and platelet engraftment. CB progenitors expanded ex vivo before transplantation provide more rapid hematopoietic and immune reconstitution as well as less engraftment failure compared with unmanipulated CB. However, the safety of infusing double and ex vivo–expanded CB has not been systematically examined.MethodsWe reviewed the immediate adverse events (AE) associated with the infusion of CB occurring within 24 hours in 137 patients enrolled in clinical CB transplant trials at the MD Anderson Cancer Center from February 2004 to May 2010. All patients received an unmanipulated CB unit followed by infusion of a second unmanipulated CB unit or a second CB unit expanded ex vivo with the use of cytokines in a liquid culture system or in mesenchymal stromal cell co-cultures.ResultsA total of three grade 2 and two grade 3 infusion reactions occurred within 24 hours of CB transplantation. This resulted in an AE rate of 3.7%. The majority of AEs manifested as signs of hypertension. No association with patient age, sex, disease status, premedication, ABO compatibility or total infusion volume was observed. In summary, the incidence of infusion-related toxicities in patients who receive unmanipulated and ex vivo–expanded double CB transplantation is low.ConclusionsWe conclude that the infusion of unmanipulated followed by expanded CB products is a safe procedure associated with a low probability of inducing severe reactions.     

Quality comparison of umbilical cord blood cryopreserved with conventional versus automated systems

Umbilical cord blood (CB) banks usually freeze and store CB for clinical transplantation using conventional controlled-rate freezer or the automated BioArchive system. The aim of this study is to compare the quality of CB cryopreserved with conventional and automated methods and to make clear the cause of the quality difference between the two methods. The experiment used 80 CB units: 40 were conventionally cryopreserved and the remainder were cryopreserved with a BioArchive. After thawing, the following measures of CB quality were compared: recovery rates of cell count, cell viability of total nucleated cells (TNCs), mononuclear cells (MNCs), and CD34+ cells, as well as colony-forming unit-granulocyte/macrophage (CFU-GM) content. Additionally, processing and storage records were reviewed to quantify the number of exposures of CB units at room temperature (transient warming event, TWE), which was analyzed in relation to CB quality. MNC and CD34+ cell viability were as follows: MNC, 78.2% ± 6.8% (conventional), 81.7% ± 7.2% (automated); CD34+ cell, 90.6% ± 6.9% (conventional), 94.7% ± 3.5% (automated). The absolute CFU-GM content per CB unit was 7.1 × 10⁵ ± 5.9 × 10⁵ with conventional cryopreservation and 12.3 × 10⁵ ± 12.0 × 10⁵ with automated cryopreservation. CBs cryopreserved with BioArchive showed significantly higher MNC and CD34+ cell viability, and CFU-GM content than those conventionally cryopreserved. The CB quality comparison depending on the amount of TWEs showed no significant quality difference between groups that were more exposed to TWEs and groups that were less exposed. CBs cryopreserved with BioArchive were of higher quality than conventionally cryopreserved CBs, and the cause of quality difference might be due to the difference of freezing conditions rather than the TWE effect.     

影响脐带血采集质量的相关性因素分析

目的回顾性分析影响脐带血采集质量的母婴因素和采集处理因素。 方法记录389份脐带血的采集量、母亲年龄、孕龄、新生儿体重、分娩方式、新生儿性别、胎次及脐带血采集至计数间隔以及采集方式。用KX-21型全自动血液分析仪进行细胞计数并计算有核细胞总数（TNC）。采用Pearson相关和Spearman秩相关进行相关性分析,采用t检验、Mann-Whitney U检验、方差分析及Kruskal-Wallis H检验进行分组比较,分析影响脐带血采集量和TNC的相关因素。? 结果脐带血采集量与TNC显著相关（r = 0.723,P < 0.001）,脐带血采集量大于80 ml时的TNC显著高于低体积者。在母婴因素中,母亲年龄与采集量及TNC差异均无统计学意义;孕龄与采集量负相关（r = -0.119,P = 0.019）,而与TNC正相关（r = 0.138,P = 0.007）,足月儿脐带血的TNC显著高于早产儿（P = 0.038）;婴儿出生体重与采集量及TNC均正相关（r = 0.236,P < 0.001;r = 0.275,P < 0.001）,体重较大婴儿脐带血的采集量和TNC均显著高于体重较小者（P?< 0.001）;剖宫产的脐带血采集量虽高于阴道分娩（P < 0.001）,但其TNC不及阴道分娩;男婴与女婴在脐带血采集量和TNC之间无显著差异。在采集处理因素中,脐带血采集至计数的时间间隔与采集量及TNC均无显著相关。 结论为提高脐带血的保存质量,应侧重选择胎儿体重较大、经阴道分娩的产妇作为脐带血供者,实验室应首先处理采集量较大的脐带血。     

Mesenchymal stem cells from cryopreserved human umbilical cord blood

Umbilical cord blood (UCB) is well known to be a rich source of hematopoietic stem cells with practical and ethical advantages, but the presence of mesenchymal stem cells (MSCs) in UCB has been disputed and it remains to be validated. In this study, we examined the ability of cryopreserved UCB harvests to produce cells with characteristics of MSCs. We were able to obtain homogeneous plastic adherent cells from the mononuclear cell fractions of cryopreserved UCB using our culture conditions. These adherent cell populations exhibited fibroblast-like morphology and typical mesenchymal-like immunophenotypes (CD73+, CD105+, and CD166+, etc.). These cells presented the self-renewal capacity and the mesenchymal cell-lineage potential to form bone, fat, and cartilage. Moreover, they expressed mRNAs of multi-lineage genes including SDF-1, NeuroD, and VEGF-R1, suggesting that the obtained cells had the multi-differentiation capacity as bone marrow-derived MSCs. These results indicate that cryopreserved human UCB fractions can be used as an alternative source of MSCs for experimental and therapeutic applications.     

脐血干细胞与创面修复

脐血干细胞是一类具有多向分化潜能的原始祖细胞,具备自我更新和增殖的能力,在特定条件诱导下可以分化为不同细胞,逐渐作为临床组织工程的来源细胞。近年来随着对脐血干细胞的不断研究,发现其在创面修复中具有明显的优势,成为创面临床治疗的一条新途径。本文从脐血干细胞的生物学特性、采集与冻存、体外扩增等方面对创面修复的研究进行综述。     

脐血干细胞与创面修复

脐血干细胞是一类具有多向分化潜能的原始祖细胞,具备自我更新和增殖的能力,在特定条件诱导下可以分化为不同细胞,逐渐作为临床组织工程的来源细胞。近年来随着对脐血干细胞的不断研究,发现其在创面修复中具有明显的优势,成为创面临床治疗的一条新途径。本文从脐血干细胞的生物学特性、采集与冻存、体外扩增等方面对创面修复的研究进行综述。     

Complete nationwide survey on umbilical cord blood freezing bag breakage in Japan

Background aimsAlthough umbilical cord blood (UCB) has now become a common stem cell source, UCB bag breakage is a known risk in UCB transplantation (UCBT). This survey provides the first comprehensive data on the frequency and causes of UCB bag breakage in Japan.MethodsData regarding UCB bag breakage from all causes, identified between April 1, 2010, and September 3, 2013, were collected from all transplant centers registered for UCBT (209 hospitals) and all public cord blood banks (CBBs) (8 CBBs) in Japan.ResultsSeventeen incidents of UCB bag breakage at CBBs were confirmed, none of which resulted in bags being shipped to transplant centers. From among 3836 UCBT, 16 incidents (0.4%) of UCB bag breakage were confirmed at transplant centers. Although all these bags were used for transplantation, no direct health hazard was reported. The major cause of UCB bag breakage confirmed at transplant centers was considered to be external force (75%). In addition, 11 incidents of unexplained UCB bag breakage at sealing between compartments were reported.ConclusionsUCB bag breakage was confirmed at both CBBs and transplant centers. UCB bags should be handled with particular care and attention.     

Quality evaluation of umbilical cord blood progenitor cells cryopreserved with a small-scale automated liquid nitrogen system

The performance of a small-scale automated cryopreservation and storage system (Mini-BioArchive system) used in the banking of umbilical cord blood (UCB) units was evaluated. After thawing the units, the viability and recovery of cells, as well as the recovery rate of hematopoietic progenitor cells (HPCs) such as CD34⁺ cells, colony-forming unit-granulocyte-macrophage (CFU-GM), and total CFU were analyzed. Twenty UCB units cryopreserved using the automated system and stored for a median of 34 days were analyzed. Mean CD34⁺ cell viabilities before freezing were 99.8 ± 0.5% and after thawing were 99.8 ± 0.4% in the large bag compartments and 99.7 ± 0.5% in the small compartments. The mean recovery values for total nucleated cells, CD34⁺ cells, CFU-GM, and total CFU were 94.8 ± 16.0%, 99.3 ± 18.6%, 103.9 ± 20.6%, and 94.3 ± 12.5%, respectively in the large compartments, and 95.8 ± 25.9%, 106.8 ± 23.9%, 101.3 ± 23.3%, and 93.8 ± 19.2%, respectively in the small compartments. A small-scale automated cryopreservation and storage system did not impair the clonogenic capacity of UCB HPCs. This cryopreservation system could provide cellular products adequate for UCB banking and HPC transplantation.     

Optimal umbilical cord blood collection,processing and cryopreservation methods for sustained public cord blood banking

Background aimsUmbilical cord blood is an established source of stem cells in patients with hematologic malignancies who do not have HLA-compatible matched related or unrelated donors. The success of an umbilical cord blood transplant depends on the dose of total nucleated and CD34+ cells infused. Therefore, collecting, banking and listing high-quality cord blood units with high total nucleated and CD34+ cell dose are essential.MethodsHere the authors describe their cord blood bank's novel collection technique, which involves both in utero and ex utero collection of a single cord blood unit. The authors also evaluated maternal, neonatal and collection parameters that may impact the cell dose.ResultsMaternal gestational age and race, and neonatal weight and sex correlated with the total nucleated cell dose.ConclusionsThe optimized collection of umbilical cord blood is critical for its use as a source of stem cells for transplantation.     

脐带血干细胞的基础与应用研究

作为造血干/祖细胞(hematopoieticstemcells/hematopoieticprogenitorcells,HSCs/HPCs)的另一来源,脐带血已经应用于临床治疗多种恶性和非恶性疾病。脐带血中HSCs/HPCs的质与量是决定其临床应用效果的最重要因素。同时,脐带血中还存在多种非造血的干细胞和前体细胞,如间充质干细胞(mesenchymalstemcells,MSCs)、内皮前体细胞(endothelialprogenitorcells,EPCs)和非限制性体干细胞(unrestrictedsomaticstemcells,USSCs)等,这些细胞可能会在未来的细胞治疗和再生医学中发挥重要作用。本综述还讨论了脐带血的临床应用及HSCs/HPCs的体外扩增、增加HSCs归巢和再植能力等提高其临床应用能力的相关研究。     

WJSC 6th Anniversary Special Issues（1）:Hematopoietic stem cell transplantation Allogeneic hematopoietic cell transplant for acute myeloid leukemia:Current state in 2013 and future directions

Acute myeloid leukemia(AML)represents a heterogeneous group of high-grade myeloid neoplasms of the elderly with variable outcomes.Though remissioninduction is an important first step in the management of AML,additional treatment strategies are essential to ensure long-term disease-free survival.Recent pivotal advances in understanding the genetics and molecular biology of AML have allowed for a risk-adapted approach in its management based on relapse-risk.Allogeneic hematopoietic cell transplantation(allo-HCT)represents an effective therapeutic strategy in AML providing the possibility of cure with potent graft-versus-leukemia reactions,with a demonstrable survival advantage in younger patients with intermediate-or poor-risk cytogenetics.Herein we review the published data regarding the role of allo-HCT in adults with AML.We searched MEDLINE/PubMed and EMBASE/Ovid.In addition,we searched reference lists of relevant articles,conference proceedings and ongoing trial databases.We discuss the role of allo-HCT in AML patients stratified by cytogenetic-and molecular-risk in first complete remission,as well as allo-HCT as an option in relapsed/refractory AML.Besides the conventional sibling and unrelated donor allografts,we review the available data and recent advances for alternative donor sources such as haploidentical grafts and umbilical cord blood.We also discuss conditioning regimens,including reduced intensity conditioning which has broadened the applicability of allo-HCT.Finally we explore recent advances and future possibilities and directions of allo-HCT in AML.Practical therapeutic recommendations have been made where possible based on available data and expert opinion.     

Analyses to clarify rich fractions in hepatic progenitor cells from human umbilical cord blood and cell fusion

Umbilical cord blood (UCB) is a source of hematopoietic stem cells and other stem cells, and human UCB cells have been reported to contain transplantable hepatic progenitor cells. However, the fractions of UCB cells in which hepatic progenitor cells are rich remain to be clarified. In the present study, first, the fractionated cells by CD34, CD38, and c-kit were transplanted via portal vein of NOD/SCID mice, and albumin mRNA expression was examined in livers at 1 and 3 months posttransplantation. At 1 and 3 months, albumin mRNA expression in CD34+UCB cells-transplanted livers was higher than that in CD34- cells-transplanted livers. Albumin mRNA expression in CD34+CD38+ cells-transplanted livers was higher than that in CD34+CD38- cells-transplanted [corrected] liver at 1 month. However, it was much higher [corrected] in CD34+CD38- cell-transplanted livers at 3 months. Similar expression of albumin mRNA was obtained between CD34+CD38+c-kit+ cells- and CD34+CD38-c-kit- cells-transplanted livers, and between CD34+CD38-c-kit+ cells- and CD34+CD38-c-kit- cells-transplanted livers, respectively. Second, fluorescence in situ hybridization and immunohistochemistry were performed to examine whether UCB cells really transdifferentiated into hepatocytes or they only fused with mouse hepatocytes. In mouse liver sections, of 1.2% cells which had human chromosomes, 0.9% cells were due to cell fusion, whereas 0.3% cells were transdifferentiated into human hepatocytes. These results suggest that CD34+UCB cells are rich fractions in hepatic progenitor cells, and that transdifferentiation from UCB cells into hepatocytes as well as cell fusion simultaneously occur in this situation.     

Expanded umbilical cord blood T cells used as donor lymphocyte infusions after umbilical cord blood transplantation

BackgroundUmbilical cord blood (UCB) is an alternative graft source for hematopoietic stem cell transplantation and has been shown to give results comparable to transplantation with other stem cell sources. Donor lymphocyte infusion (DLI) is an effective treatment for relapsed malignancies after hematopoietic stem cell transplantation. However, DLI is not available after UCB transplantation.MethodsIn this study, in vitro–cultured T cells from the UCB graft were explored as an alternative to conventional DLI. The main aim was to study the safety of the cultured UCB T cells used as DLI because such cell preparations have not been used in this context previously. We also assessed potential benefits of the treatment.ResultsThe cultured UCB T cells (UCB DLI) were given to 4 patients with mixed chimerism (n = 2), minimal residual disease (n = 1) and graft failure (n = 1). No adverse reactions were seen at transfusion. Three of the patients did not show any signs of graft-versus-host disease (GVHD) after UCB DLI, but GVHD could not be excluded in the last patient. In the patient with minimal residual disease treated with UCB DLI, the malignant cell clone was detectable shortly before infusion but undetectable at treatment and for 3 months after infusion. In 1 patient with mixed chimerism, the percentage of recipient cells decreased in temporal association with UCB DLI treatment.ConclusionsWe saw no certain adverse effects of treatment with UCB DLI. Events that could indicate possible benefits were seen but with no certain causal association with the treatment.