脐血采集方式对脐血造血细胞含量的影响

目的比较阴道分娩和剖宫产采集脐血所含造血细胞含量的差异,评估两种采集方式的优缺点。方法回顾性分析两种脐血采集方式间新生儿体重、采集体积、总有核细胞数(TNC)、CD34+细胞百分率、CD34+细胞数和总集落数(CFUs)的差异。结果采集体积在两组间差异无统计学意义(P>0.05)。而阴道分娩组TNC、CD34+细胞百分率、总CD34+细胞数、CFUs数明显高于剖宫产组,新生儿体重小于剖宫产组,差异有统计学意义(P<0.05)。新生儿体重在2.5kg～4.0kg时,阴道分娩组TNC、总CD34+细胞数和CFUs数明显高于剖宫产组,差异有统计学意义(P<0.05)。结论阴道分娩采集方式采集的脐血含有较高的造血细胞含量且有利于采集到更好脐血质量,可作为同胞脐血移植采集脐血的最优选分娩和采集方式。     

母婴因素对脐血CD34+造血干/祖细胞的影响

目的 比较不同母婴因素与脐血有核细胞总数及CD34⁺造血干/祖细胞数量的关系,为脐血库合理选择脐血提供参考。方法 前瞻性收集130例2019年6月至2020年1月期间于大连市妇女儿童医疗中心分娩的新生儿脐血标本,男女比例为1：1。收集围生期相关信息：孕母年龄及血型,有/无妊娠糖尿病、妊娠高血压,妊娠方式、分娩方式、单胎/双胎,新生儿体重、性别、生后Apgar评分,以及胎盘、羊水、脐带情况。结果 根据孕母血型、妊娠糖尿病、妊娠高血压、妊娠方式、分娩方式、单胎/双胎,新生儿性别、生后Apgar评分,胎盘形态、羊水胎粪污染、脐带绕颈等情况进行分组,组间比较脐血有核细胞总数及CD34⁺细胞计数差异均无统计学意义（P > 0.05）。孕母年龄、新生儿体重与脐血有核细胞总数无相关性（P > 0.05）,新生儿体重与CD34⁺细胞计数无相关性（P > 0.05）,孕母年龄与CD34⁺细胞计数呈正相关（P < 0.05）。结论 脐血中CD34⁺细胞数量随孕母年龄增大而增多,故脐血库在筛选脐血时,同等条件下可以选择年龄偏大的孕妇。     

脐血造血细胞含量与白血病脐血移植疗效分析

目的分析4711份库存脐血造血细胞含量及探讨脐血造血细胞含量与白血病脐血移植疗效的关系。方法分析4711例库存脐血总有核细胞数(TNC)和CD34+细胞数分布情况,探讨不同的造血细胞输入量、供受者HLA不相合数、受者性别、年龄、体重和疾病类型间植入率和生存率的差异。结果 4711例库存脐血TNC和CD34+细胞中位数分别为1.14×109/kg和4.06×106/kg,按3.7×107/kg有效TNC输入量计算,93.2%脐血可供体重50 kg以下受者移植。89例白血病患者移植后植入75例,植入率为84.3%。中性粒细胞绝对值≥0.5×109/L、血小板≥20×109/L和≥50×109/L的时间分别为移植后17、34和46 d。75例植入病例中,长期无病存活47例,死亡26例,2例复发;急性移植物抗宿主病(GVHD)Ⅰ～Ⅱ度、Ⅲ～Ⅳ度和慢性GVHD发生率分别为54.7%、20.0%、9.3%。影响移植植入率的因素包括受者年龄、TNC和CD34+细胞输入量;影响生存率的因素包括受者年龄、体重和输入CD34+细胞数。结论在无法找到HLA全相合骨髓供者时,可选择脐血作为替代骨髓的造血干细胞来源治疗儿童与成人白血病,TNC和CD34+细胞数仍是选择脐血移植物的参考指标。     

群体反应性抗体干扰脐血CD34+细胞增殖、分化的实验研究

目的 探讨血清特异性群体反应件抗体(PRA)对脐血CD34+细胞增殖、分化能力的影响.方法 取含PRA(经实验证实)的β地中海贫血患儿血清,与脐血CD34+细胞、补体孵育,观察PRA对CD34+细胞增殖、分化的影响,分A组(不加血清组)、B组(PRA血清组)、C组(PRA血清加补体组)、D组(补体组)、E组(PRA阴性血清组)共5组.孵育后以3H-TaR掺入法测定细胞DNA合成及流式细胞仪检测Annexin V和CD95表达;并进行集落培养,于第10天计数集落.结果 A组为(20.71±2.81)U/L,低于B组(64.28±5.12)U/L、C组(84.29±4.99)U/L,B组低于C组;D、E组均为(22.86±2.91)U/L和(22.86±2.91)U/L,均低于B、C组;各组氚每分钟β射线释放量(cpm):A组为(22629±3288),高于B组(4598±2178)和C组(1626±1192),A组和D、E组之间的差异无统计学意义(P＞0.05);A组的总集落数、粒-巨噬细胞集落形成单位(CFU-GM)、混合系集落形成单位(CFU-GEMM)及爆式红系集落形成单位(BFU-E)数均高于B、C组,B组的总集落数、CFU-GM及CFU-GEMM数均高于C组;D组和E组的各种集落数与A组的差异无统计学意义(P＞0.05);各组CD34+细胞Annexin V及CD95表达百分率差异无统计学意义(P＞0.05).结论 特异件PRA血清对脐血CD34+细胞的增殖和分化有抑制作用,补体可增强上述作用;特异性PRA血清对脐血CD34+细胞的凋亡无明显影响.     

早产儿脐血造血干/祖细胞特点

目的对12例早产儿和18例足月儿脐血采集量、有核细胞数量、CD34+细胞比例、体外形成造血细胞克隆的能力进行比较研究。方法采用密闭式脐血采集袋,经脐静脉穿刺收集脐带血,按体积比5:1与60 g.L-1羟乙基淀粉(HES)混合、浓缩有核细胞,流式细胞仪检测CD34+细胞比例,甲基纤维素半固体培养基检测其脐血形成造血细胞克隆能力。结果早产儿脐血采集量及分离前后有核细胞数量均低于足月儿脐血[脐血采集量分别为(76.52±22.48)mLvs(94.21±20.32)mL,P<0.05;分离前有核细胞数量分别为(4.78±2.30)×108vs(8.36±2.51)×108,P<0.01;分离后有核细胞数量分别为(3.72±1.71)×108vs(6.54±1.94)×108,P<0.01)]。早产儿CD34+细胞比例高于足月儿脐血[(0.49±0.16)%vs(0.32±0.13)%,P<0.01],早产儿脐血体外生成红系爆式集落形成单位和粒-单系集落形成单位的能力也高于足月儿脐血[分别为(29.58±10.54)/2×105MNCvs(19.27±7.26)/2×105MNC,P<0.01和(45.28±16.2...     

脐血的采集和分离及有核细胞的富集

脐血中富含造血干 /祖细胞 ,是骨髓和外周血后的第三种造血干细胞的来源[1] 。脐血的采集、分离和保存方式对单个核细胞 (ＭＮＣ)和ＣＤ 3 4细胞的回收有一定的影响。分析不同性别、胎龄、胎次和体重的胎儿脐血量、ＭＮＣ数 ,以及脐血采集后处理方式对ＭＮＣ和ＣＤ 3 4细胞的影响 ,有助于脐血临床移植和建立脐血库。材料和方法1.标本来源 :所有 83例脐血均来自本院产科 ,其中男性 48例 ,女性 35例。胎龄最小为 30周 ,最大 42周 ,平均体重 (3 4± 0 4)ｋｇ ,剖宫产 36例 ,自然分娩 47例。2 .脐血采集 :采用开放式采集。在胎儿娩出后 5ｍｉ…     

人脐血间充质干细胞对脐血CD+34细胞体外扩增作用的研究

目的 探讨含人脐血来源的间充质干细胞(MSCs)体系在体外对脐血造血干细胞(HSCs)扩增作用。方法 (1)用含人脐血MSCs及不同造血生长因子(HGFs)组合的无血清扩增体系对人脐血CD34^ 细胞进行体外扩增。(2)于扩增前及扩增后第6、12天分别用双色流式细胞仪动态检测HSCs表面抗原标记：CD34^ 、CD34^ CD38^-、CD34^ CD3^ 、CD34^ CD19^ 、CD34^ 和CD34^ CD41^ 。细胞的含量。(3)按本实验室方法行体外半固体培养,观察扩增前后脐血细胞粒-单核细胞集落形成单位(CFU-GM)、爆式红系集落形成单位(BFU-E)、混合集落形成单位(CFU-Mix)及高增殖集落形成单位(CFU-HPP)集落形成情况。结果(1)含人脐血MSCs体系对脐血CD34^ CD38^ 细胞的扩增倍数在第6天和第12大分别为159和437倍。该业群百分比在单纯因子组扩增第12天时为1．98％,而在含脐血MSCs组为9．98%,明显高于扩增前。(2)集落培养表明,含脐血MSCs组扩增第12天与扩增第6天相比,其CFU-Mix和CFU-HPP的扩增倍数增加,而单纯因子组这两种集落的扩增倍数下降。(3)随扩增天数的增加,两组扩增体系中CD34^ CD3^ 和CD34^ CD41a^ 细胞均明显增加,而CD34^ CD19^ 和CD34^ CD3^ 细胞均明显减少。两组相比,含脐血MSCs组差异更显著。结论 (1)含脐血MSCs体系不仅能扩增更原始的造血干／祖细胞(HSPC),且具有在短期内(12d)保持HSCs不耗竭。(2)含脐血MSCs体系对脐血CD34^ 细胞向定向祖细胞的扩增,主要为髓系及巨核系祖细胞,而对其向淋巴系祖细胞的扩增具有抑制作用。     

肿瘤患儿外周血单个核细胞采集及其对受体造血恢复的影响

目的回顾分析26例白血病和实体瘤患儿外周血干细胞采集效果及其对受体造血恢复影响,筛选可能与采集效率提高和受体造血恢复相关的因素。方法总结26例供体的49次粒系集落刺激因子(G-CSF)动员后的外周血采集过程,统计分析供体动员剂量、采集时外周血白细胞、收获的单个核细胞(MNC)和CD34 细胞数、受体造血恢复三者间的相关性。结果26例供体均能很好耐受动员和采集过程。采集物MNC和CD34 细胞中位数分别为4.5×108/kg和1.9×106/kg。动员剂量与采集物中MNC数呈正相关,与采集物CD34 细胞数无相关。采集当天供体外周血白细胞总数对采集细胞量无影响,而MNC水平与CD34 细胞采量正相关。结论受体的造血恢复与采集物中的CD34 细胞数可能存在相关性。因此这类患儿的采集时有必要动态监测其动员时的外周血CD34 细胞水平,确定最佳采集时间,以获取足够的CD34 细胞采量。     

足月新生儿脐血血清游离卡尼汀浓度及相关因素研究

目的了解足月新生儿脐血血清游离卡尼汀浓度及其相关影响因素。方法收集2004年11月至2005年2月出生的足月新生儿89例,男46例,女43例。平均胎龄(39.4±0.9)周,出生体重(3427.2±414.4)g,身长(49.8±2.5)cm,头围(34.8±1.4)cm,Apgar评分为8~10分。平产50例,剖宫产39例。于出生后即采集脐血2ml,用高效液相(HPLC)法测定脐血血清中卡尼汀浓度。结果89例正常新生儿平均脐血血清卡尼汀浓度为(52.2±22.9)μmol/L。其中男性组为(49.3±18.7)μmol/L,女性组为(55.3±26.5)μmol/L,两者之间差异无显著性(P=0.216);平产组为(52.4±24.5)μmol/L,剖宫产组为(51.9±20.9)μmol/L,两者之间差异无显著性(P=0.924)。根据新生儿出生体重分为<3000g(11例)、3000~3999g(68例)和≥4000g(10例)3组,其血清卡尼汀平均值分别为(61.5±26.2)μmol/L、(52.0±23.1)μmol/L和(42.8±12.3)μmol/L。脐血卡尼汀浓度与出生体重呈负相关(r=-0.239,P=0.026);与孕母年龄、新生儿胎龄、身长和头围无相关性。结论本组足月新生儿脐血血清卡尼汀浓度为(52.2±22.9)μmol/L,与新生儿出生体重呈负相关,与孕母年龄、新生儿胎龄、身长、头围、性别和分娩方式无相关性。     

新生儿脐血pH和BE影响因素的研究

目的通过大样本收集新生儿脐血血气,研究脐血血气统计学参考值范围与不同影响因素的相关关系。方法选择2012年5～11月广东省妇幼保健院和新会妇幼保健院产科出生的新生儿进行前瞻性研究,选取其中1rainApgar评分〉7分者的脐血血气结果进行统计分析,了解正常新生儿脐血血气的统计学参考值范围;重点分析影响新生儿脐血pH和BE的因素。结果2000例新生儿中,1min Apgar评分≤7分11例,〉7分1989例,低Apgar评分组pH〈7．2的比例为45．5％,正常Apgar评分组pH〈7．2的比例为3．5％,差异有统计学意义（P〈0．001）;1800例足月单胎、体重适于或大于胎龄新生儿中,1794例1min Apgar评分〉7分者脐血pH和BE的统计学参考值范围分别是7．34±0．14（X±1．96S）和-3．53±6．57（X±1．96s）。单因素分析显示,宫内窘迫组、妊娠期并发症组pH值均低于对照组,剖宫产组pH和BE值均高于阴道分娩组,脐带绕颈组pH值降低,双胎组BE值高于单胎组;羊水性状对pH、BE值均无影响。多因素分析显示,宫内窘迫、分娩方式均对脐血血气有影响。结论足月单胎、体重适于或大于胎龄新生儿中,1min Apgar评分〉7分者脐血pH值和BE值的统计学参考值范围分别是7．34±0．14和-3．53±6．57;Apgar评分与脐血血气分析具有一致性,但单独使用Apgar评分诊断早产儿窒息可能会增加窒息的误诊率;宫内窘迫可能会增加新生儿酸中毒的发生率,不同分娩方式对脐血血气pH、BE值均有影响。     

Impact of maternal and neonatal factors on CD34+ cell count, total nucleated cells, and volume of cord blood

The engraftment outcome of UCB transplantation is highly dependent on cell number. It would be useful to predict CB cell content using information of donor-related variables before cell processing. In this study, CBs were obtained from 1312 single-birth term deliveries in the Buddhist Tzu Chi Stem Cells Center from January 2001 to June 2006. We evaluated whether maternal factors, such as age and race, have an effect on laboratory parameters of hematopoietic content, including CD34+ cell counts, TNCs, and cord blood volume. We also studied the impact of neonatal factors, such as delivery method, gestational age, sex, birth weight, and birth order on the same parameters. In multivariate analysis, babies delivered via Cesarean section had more CD34+ cells and volume, but lower TNCs. Similar results were found for either babies of shorter gestational age or in male infants. Babies with larger birth weight had higher CD34+ cell volume, and TNC, while mothers with fewer previous live births had CB with more TNCs. Maternal age and race had no effect on these laboratory parameters. To conclude, our results suggest that neonatal factors affect CB cell yields. TNCs tend to be more affected by different variables than CD34+ cell counts and volume. These findings may help in collecting CB efficiently and improve the CB transplantation rate.     

Does umbilical cord blood-derived CD34+ cell concentration depend on the weight and sex of a full-term infant?

Umbilical cord blood has recently been considered as an alternative source of hematopoietic progenitor cells for clinical application. Patient survival in allogenic cord blood transplantation is critically dependent on the cord blood-derived total nucleated cell count and the total CD34 cell count/kg of the body weight. A number of factors such as maternal age, gestational age, newborn's sex and weight, umbilical cord length, and placental weight can influence the volume, amount of mononuclear cells, and the CD34 cell concentration. Cord blood was collected from normal vaginal and cesarean deliveries. It was immediately processed and assessed for the total nucleated cell count and CD34 cell concentration. Assessment of maternal and neonatal parameters such as gestational age, baby's birth weight, and sex was carried out with the CD34 cell concentration. The mean CD34 cell concentration was 0.21±0.24% for the group with <2500 g (low birth weight) birth weight of the baby (n=104). The mean CD34 cell concentration was 1.84±1.12% for the group with ≥2500 g (normal birth weight) birth weight of the baby (n=396). A strong positive correlation was found between birth weight of the baby with cord blood-derived CD34 cell concentration (*P<0.0001, r=0.81). A positive correlation was found between gestational age and cord blood-derived CD34 cell concentration (*P<0.0001, r=0.31). No significant correlation was found between the baby's sex with the CD34 cell concentration and the total nucleated cell count. This study concludes that the higher the birth weight of the baby, the better the yield of the CD34 cell concentration, and hence these should be the preferred samples for infusion.     

Serum zinc and copper levels in the maternal blood and cord blood of neonates

Estimation of serum zinc and copper in the maternal blood and cord blood of neonates was carried out to correlate the trace metals in the neonates and their mothers in relation to gestational age and birth weight. Sixty-five healthy neonates, both term and preterm and their mothers were selected. This cross sectional study was done at Azimpur Maternity Centre, Dhaka Medical College Hospital and Chemistry Division, Atomic Energy Centre, Dhaka, Bangladesh from July 1997 to June 1998. The estimation of trace metals was carried out by Atomic Absorption Spectrophotometry (AAS). The mean serum zinc levels in the maternal blood and cord blood were 0.47 ± 0.24 μg/ml and 0.85 ± 0.33 μg/ml respectively and the mean copper levels in the maternal blood and cord blood were 1.37 ± 0.62 μg/ml and 0.31 ± 0.32 μg/ml respectively. Cord blood zinc level was significantly higher and cord blood copper level was significantly lower than the corresponding maternal blood levels. There was no significant correlation between gestational age and serum zinc levels in the cord or maternal blood. But significant inverse correlation was found between gestational age and serum levels of copper in the maternal and cord blood.     

Immunophenotypic changes of fetal cord blood hematopoietic progenitor cells during gestation

We measured cell surface expression of CD34, HLA-DR, CD38, CD19, CD33, CD71, and CD45 antigens in the hematopoietic progenitor cells of fetal cord blood to investigate immunophenotypic changes at different gestational ages. These antigens were identified by flow cytometry in 11 fetuses (gestational age 19-24 wk, in 12 preterm (25-28 wk) and in ten newborn infants born at term. The frequency and number of CD34+ cells were higher in the blood of the 11 fetuses; in addition, a statistically significant inverse correlation between number of CD34+ cells and advancing gestational age was noted. The numbers of CD34+ CD19+, CD34+ CD33+, and CD34+ CD45+ coexpressing cells were significantly higher in the fetuses, whereas CD34+ CD38+ cells were more represented in the neonates at term. Gestational age was inversely correlated with the number of CD34+ CD19+ and CD34+ CD33+ coexpressing cells. A positive correlation between gestational age and CD34+ CD38+ cells was noted. The number of CD34- CD19+, CD34- CD38+, and CD34- CD45+ cells was higher in term infants; furthermore, a significant correlation between advancing gestational age and CD34- CD38+ or CD34- CD45+ cells was demonstrated. The proliferative capacity was also higher at lower gestational ages. These data suggest that the development and lineage commitment of fetal cord blood hematopoietic progenitor cells are very active during the last two trimesters of pregnancy. The most significant changes of hematopoietic cells maturation seem to occur within 25 wk of gestation.     

Cord blood erythropoietin in the presence and absence of labor in normal infants

To evaluate erythropoietin (Ep) levels in normal labor and cesarean section we studied the cord serum of 111 term pregnancies, divided into three groups: (i) those born by normal vaginal delivery (n = 69); (ii) those delivered vaginally from mothers who were smokers (n = 20); and (iii) those delivered by elective cesarean section (n = 22). The three groups did not differ in maternal age, gestational age, birthweight, infant sex and Apgar scores. No correlation was found between Ep and hematocrit in all three groups of normal terms studied. Although not statistically significant the lower mean Ep value observed in cesarean section compared with the values obtained from normal deliveries could suggest that the process of labor may be a cause of these differences.     

Birthweight of full-term infants is associated with cord blood CD34+ cell concentration

AIM: CD34+ cell counts are used to define the haematopoietic stem cell potential of a given cord blood transplant. The aim was to test the hypothesis that high concentration of cord blood haematopoietic progenitor and stem cells could be a reflection of intrauterine growth, of which birthweight is an indicator. METHODS: Simple and multiple regression analyses were applied to test cord blood bank data on 1368 infants for associations of selected obstetric factors and cellular contents of cord blood. RESULTS: When groups were formed based on the extreme values (5th versus 95th percentiles) of a given variable, e.g. birthweight, the term infants having the highest birthweights were found to have statistically significantly higher median cord blood CD34+ cell concentrations. Also, infants in the top 50th percentile of relative birthweight had higher median CD34+ cell concentration than infants in the low 50th percentile. In multiple regression analysis, the correlation between birthweight and CD34+ cell concentration was statistically clearly significant. Notably, while an expected correlation between gestational age and nucleated cell concentration was found, there was no association between infant gestational age and CD34+ cell concentration. CONCLUSION: Haematopoietic progenitor and stem cells may reflect intrauterine growth and have a more central role in foetal development than has been reported earlier.     

Interleukin-10 and transforming growth factor-β1 in cord blood: relationship with paternal allergy and cesarean section

Aim: To measure Interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) in cord blood and assess their relationship with parental allergy and perinatal characteristics.
Methods: In a neonatal care unit 212 consecutive full-term and appropriate for gestational age newborns were recruited. IL-10 and TGF-β1 levels were determined in cord blood by high sensitivity ELISA. Perinatal characteristics, mode of delivery and presence of allergy in parents were recorded.
Results: Out of 212 newborns, 136 were of non-allergic parents and 76 (35.8%) of one or both allergic parents. In newborns of allergic fathers median IL-10 levels tended to be lower (0.67 vs. 1.06 pg/mL, p = 0.07) and TGF-β1 levels were significantly lower (40.9 vs. 45.3 ng/mL, p = 0.008) than in newborns of non-allergic parents. Multiple general regression analysis showed that presence of paternal allergy (β=−0.19, p = 0.003) to be born by cesarean section (β=−0.21, p = 0.03) and younger gestational age (β= 0.14, p = 0.04) independently contributed to decrease TGF-β1 levels (multiple R = 0.38, p < 0.0001).
Conclusion: Paternal allergy and cesarean section are associated to decreased TGF-β1, which might be the mediator of the increased risk of atopy development. Cord blood IL-10 and TGF-β1 levels of our newborn series could be used as reference values for further studies on these relationships.