脐血巨核系造血细胞的初步研究

用甲基纤维素法培养１５例新生儿脐血巨核系造血祖细胞（ＣＦＵ－Ｍｅｇ），观察了ＣＦＵ－Ｍｅｇ的生长特性、集落产率与接种细胞数的相关性，并与成人骨髓及外周血ＣＦＵ－Ｍｅｇ的数量进行了比较。结果表明，脐血中富含ＣＦＵ－Ｍｅｇ，且分裂快、子代细胞多，形成集落早呈现活跃的增殖状态，其数量低于成人骨髓但个体间差异明显，２６．７％的样本ＣＦ－Ｍｅｇ产率高于骨髓。接种细胞数量与ＣＦＵ－Ｍｅｇ的产率在一定范围内呈良     

脐血混合移植的基础和临床研究

用单层琼脂法进行单一和混合脐血粒－单造血祖细胞（ＣＦＵ－ＧＭ）培养，结果发现不同脐血单个核细胞（ＭＮＣ）间可能互为造血细胞集落刺激活性，混合培养ＣＦＵ一ＧＭ产率明显高于单独培养之和的均值（Ｐ＜０．０５）。长期深低温保存可累积脐血造血祖细胞数量，且可降低Ｔ４淋巴细胞的抗原表达，有利减轻移植排斥。在实验基础上，对２例晚期盆腔肿瘤患儿进行了强力化疗和混合脐血移植术，病情均于一个月内完全缓解，骨髓造血重建，为造血组织移植开拓了新的供源，可能使更多的不治之症患者获得治疗的机会。     

脐血中树突状细胞的体外诱导和抗肿瘤效应的实验研究

探讨脐血来源的树突状细胞（ＤＣ）的体外诱导及其在抗肿瘤免疫中的作用。方法无菌条件下常规要脐血,采用淋巴细胞分离液分离脐血单个核细胞,去除悬浮细胞后获得单核细胞。将获得的单核细胞分为两组,实验组１在含有细胞因子白介素４（ＩＬ－４）、粒单细胞集落刺激因子（ＧＭ－ＣＳＦ）和肿瘤坏死因子（ＴＮＦ－α）的综合成培养液ＤＭＥＭ中培养,对照组１培养液中未加上述细胞因子,第１０天收集部分细胞进行细胞表型分析,并     

细胞因子在妇科恶性肿瘤化疗中的应用

本文综述了粒细胞－集落刺激因子（Ｇ－ＣＳＦ）、单核细胞系－集落刺激因子（Ｍ－ＣＳＦ）、粒细胞单核细胞系－集落刺激因子（ＧＭ－ＣＳＦ）、白细胞介素－６（ＩＬ－６）和红细胞生成素等细胞因子的组织来源、生理特性。阐述了应用其预防／治疗妇科恶性肿瘤化疗引起的骨髓抑制的方法、效果评价及本身的毒副作用。     

应用AnnexinV—FITC／PI—FCM定量分析对5—FU诱导卵巢癌 …

目的 应用ＡｎｎｅｘｉｎＶ－ＦＩＴＣ／ＰＩ－ＦＣＭ定量分析５－氟尿嘧啶（５－ＦＵ）诱导卵巢癌细胞系ＯＶＣＡＲ发生的凋亡。方法 将５－ＦＵ以０、２０ｍｇ／Ｌ、１００ｍｇ／Ｌ、２００ｍｇ／Ｌ的浓度，分别处理ＯＶＣＡＲ细胞１２、２４、４８小时，然后用形态学观察、ＤＮＡ琼脂糖电泳和ＡｎｎｅｘｉｎＶ－ＦＩＴＣ／ＰＩ流式细胞技术（ｆｌｏｗ ｃｙｔｏｍｅｔｒｉｃ，ＦＣＭ）检测其诱发调亡的情况。结果 Ａｎｎｅｘｉ     

自泌性转化生长因子－β对人卵巢癌细胞生长的影响

目的：我们以往观察到，人卵巢癌细胞系ＣＯＣ１和ＣＯＣ２在无血清培养条件下，可以分泌转化生长因子－β（ＴＧＦ－β）类似物，本研究进一步观察该ＴＧＦ－β类似物对自身细胞生长的作用。方法：分别制备ＣＯＣ１和ＣＯＣ２ＲＰＭＩ１６４０无血清培养条件的培养液ＳＦＣＭ１和ＳＦＣＭ２；观察ＳＦＣＭ１和ＳＦＣＭ２对体外培养和接种ＢＡＬＢ／ｃ裸鼠ＣＯＣ１和ＣＯＣ２细胞生长的影响，并与ＴＧＦ－β标准品相比较。结果：ＳＦＣＭ１和ＳＦＣＭ２对体外培养的自身细胞ＣＯＣ１和ＣＯＣ２均表现为剂量依赖性促进作用，ＳＦＣＭ２明显促进ＣＯＣ２细胞在裸鼠体内生长。放射性受体分析结果显示，ＣＯＣ１和ＣＯＣ２细胞均表达ＴＧＦ－β受体（结合部位）；上述ＳＦＣＭ１和ＳＦＣＭ２的促进作用均可以被抗ＴＧＦ－β中和抗体部分阻断。结论：在ＣＯＣ１和ＣＯＣ２细胞存在ＴＧＦ－β自分泌环调节机制，该机制可能与ＣＯＣ１和ＣＯＣ２细胞自主性无止境生长有关。     

固定式Cu-IUD和固定式含吲哚美辛Cu-IUD对兔宫腔冲洗液及子宫内膜基质金属蛋白酶影响的初步研究

本文应用酶谱法,观察了正常成年家兔及置固定式ＣｕＩＵＤ（ＦＣｕ－ＩＵＤ）或固定式含吲 哚美辛ＣｕＩＵＤ（ＦＩＣｕ－ＩＵＤ）的成年家兔宫腔冲洗液和子宫内膜组织中ＭＭＰｓ酶谱及其活 性变化。结果显示：ＦＣｕ－ＩＵＤ组与对照组比较,酶活性增强及酶种类明显增多。ＦＩＣｕ－ＩＵＤ 组与ＦＣｕ－ＩＵＤ组比较,部分ＭＭＰｓ酶活性明显降低,而且宫腔冲洗液和内膜组织活性降 低的酶种类有所不同。结果提示ＩＵＤ可以前列腺素为中介促进子宫局部 ＭＭＰｓ-生成,这 一作用可能与ＩＵＤ引起的异常出血有关。吲哚美辛减少ＩＵＤ引起异常出血的作用可能 与ＭＭＰｓ的生成减少有关。     

同步序贯Ⅱ治疗难治性恶性滋养细胞肿瘤20例报告

同步序贯Ⅱ治疗难治性恶性滋养细胞肿瘤２０例报告戴淑真，崔竹梅，叶元华，陈竹香，刘尔珍（青岛医学院附属医院）１９８８至１９９４年我们对２０例常规化疗（ＶＣＲ＋５－ＦＵ＋ＫＳＭ＋ＣＴＸ即同步序贯Ｉ或５－ＦＵ＋ＫＳＭ）无效或复发或耐药以及外院经不正规化疗转...     

甾体激素和血管活性物质ET／NO对培养的人子宫动脉平滑肌细胞增…

利用体外培养系统，通过＾３Ｈ－ＴｄＲ掺入和ＭＴＴ法观察了甾体激素和血管活性物质内皮素（ＥＴ）与一氧化氮（ＮＯ）在培养的人子宫动脉平滑肌细胞（ＨＵＡＳＭＣ）增殖中的作用。结果：雌激素对ＨＵＡＳＭＣ的ＤＮＡ合成和细胞增殖无影响。孕激素和ＥＴ－１可促进ＨＵＡＳＭＣ ＤＮＡ合成和细胞增殖，其作用呈剂呈依赖性。孕激素可明显增强ＨＵＡＳＭＣ对ＥＴ－１的反应，ＮＯ供体硝普钠（ＳＮＰ）抑制ＨＵＡＳＭＣ ＤＮＡ合成     

人卵巢癌细胞分泌转化生长因子－β对外周血单个核细胞增殖的影响

观察人卵巢癌细胞株ＣＯＣ＿１和ＣＯＣ＿２来源的患者癌性腹水（ＡＳ＿１，ＡＳ＿２）、癌细胞无血清条件培养液（ＳＦＣＭ＿１，ＳＦＣＭ＿２）及荷瘤裸鼠血清（ＮＳ＿１，ＮＳ＿２）对植物血凝素（ＰＩＩＡ）和白细胞介素－２（ＩＬ－２）诱导的外周血单个核细胞（ＰＢＭＣ）增殖的作用，及与转化生长因子－β（ＴＧＦ－β）的关系。结果：ＡＳ＿１和ＡＳ＿２对ＰＢＭＣ增殖有剂量依赖性抑制作用。ＳＦＣＭ和ＮＳ也具有明显抑制ＰＢＭＣ增殖的作用；但在浓度很低时，ＳＦＣＭ＿２对ＰＨＡ诱导，ＳＦＣＭ＿１、ＮＳ＿１和ＮＳ＿２对ＩＬ－２诱导的ＰＢＭＣ增殖，具有一定促进作用。上述作用特点与ＴＧＦ－β作用相似，并可被抗ＴＧＦ－β中和抗体部分阻断。提示：人卵巢癌细胞可通过产生ＴＧＦ－β，抑制宿主免疫。     

Association of stress during delivery with increased numbers of nucleated cells and hematopoietic progenitor cells in umbilical cord blood

OBJECTIVE: Umbilical cord blood can be used as a source of bone marrow repopulating cells for allogeneic stem cell transplantation. Large variations in the frequencies of white blood cells and hematopoietic progenitor cells have been found for umbilical cord blood. These variations may be due in part to specific circumstances during labor and delivery. STUDY DESIGN: In this study we analyzed the relationship between stress factors occurring during parturition and the frequencies of nucleated cells, leukocyte subsets, CD34(+) cells, and hematopoietic progenitor cells, as determined in semisolid medium cultures of umbilical cord blood. RESULTS: We observed that a prolonged first stage of labor resulted in increases in the numbers of nucleated cells, granulocytes, CD34(+) cells, and hematopoietic progenitor cells in umbilical cord blood. Evaluation of parameters that indicate stress of the infant during delivery demonstrated higher numbers of nucleated cells, granulocytes, CD34(+) cells, and hematopoietic progenitor cells in umbilical cord blood from children with lower venous pH. CONCLUSION: Longer duration stress during delivery increased the numbers of nucleated cells, granulocytes, CD34(+) cells, and hematopoietic progenitor cells, possibly by causing mobilization of various cell populations by endogenous cytokines. As long as umbilical cord blood harvesting does not interfere with the delivery, umbilical cord blood collected after stressful deliveries may provide optimal units for hematopoietic stem cell transplantation.     

Human umbilical cord blood banking and transplantation: a state of the art

Human umbilical cord blood has proven to be a feasible alternative source of hematopoietic stem cells for pediatric and some adult patients with major hematologic disorders. This has promoted the establishment of cord blood banks for use in unrelated transplants worldwide. The banking of umbilical cord blood offers many advantages: absence of donor risk, absence of donor attrition, immediate availability, and the ability to expand available donor pools in targeted ethnic and racial minorities currently underrepresented in all bone marrow registries. Preliminary clinical experience suggests that, due to the immunological immaturity of cord blood cells, graft versus host disease might be lower than when using bone marrow from adult donors and HLA restrictions might be less stringent. Techniques to improve the efficacy of blood banks are currently under investigation. Closed cord blood collection methods have proven to be superior to open in reducing the risk of microbial contamination. Efficient banking requires volume reduction of cord blood units without significant loss of progenitor cells, in order to decrease storage space and cost, and this may be achieved by using the separation techniques. Cryopreservation and thawing techniques have been established and do not seem to affect the viability and progenitor cell recovery or the feasibility of CD34(+) selection and ex vivo expansion. Nevertheless, many scientific, ethical, and social questions have arisen in connection with cord blood banking that need to be addressed.     

Umbilical Cord Blood Banking: A Current Perspective

Placental and umbilical cord blood (UCB) is becoming an important source of haematopoietic stem cells for use in clinical transplantation. Now that over 500 cord blood transplantations have occurred worldwide, clinical experience is demonstrating some distinct advantages to these cells over the traditional bone marrow sources. These advantages include distinctive proliferative capacities which favour engraftment. Umbilical cord blood use is associated with a reduced incidence and severity of Graft versus Host disease, and recipients may tolerate a greater number of HLA mismatches than with bone marrow source stem cells.Umbilical cord blood stem cells have also opened up areas of active laboratory and clinical investigation that could ultimately result in further therapeutic options. The most exciting areas are those of ex vivo cell expansion and a variety of applications involving stem cells as vehicles for potential gene therapy.Cord blood banks can retain these separated stem and progenitor cells to provide an additional, immediately accessible resource to a bone marrow registry. In Europe, NETCORD links three cord blood banks to its bone marrow donor worldwide registry, and in the United States, three cord blood banks are also linked to a central bone transplant registry. In Canada, the Bone Marrow Transplant Registry, administered by The Canadian Red Cross, is investigating options for a similar structure. However, there is no established funding at this time for a Canadian system of this scale.     

The in vivo study of myeloprotection by GST-π gene transfected human cord blood hematopoietic stem cells transplantation

To investigate the influence of GST-π gene transfer into human cord blood hematopoietie stem cells on their drug resistance against anti-turmor drugs in vivo. Methods: GST-π gene transfeetion into human cord blood CD34 ceils was carried out using a retrovirus vector PTJ-GST-π with the aid of fibronectin. Successful gene transfer was confmncd by in vitro colonyassay and RT-PCR. GST-π gene transduced human cord blood CD34 ceils were then engrafted into4-week-old to tal body irradiated NOD/Scid mice and earboplatin was intraperitoneally administered sequentially at 4 weeks interval 4 weeks after engraftment.Results:Peripheral bloed(PB) WBC wassignificantly higher in GST-π mice than control mice after 2 course of carboplatin. Retroviral GST-π expression in bone marrow hematopoietie progenitor cells of recipient mice was detected by RT-PCR16 weeks after Xenotransp/antatlon. Conclusion: The transfect/on of GST-π gene could confer, tosome extent, reslstance to cord blood stem cells against carboplatin in vivo.     

Highly efficient gene transfer into preterm CD34 hematopoietic progenitor cells

OBJECTIVE: Retrovirus-mediated gene transfer has been shown to transduce CD34(+) cells from term gestation umbilical cord blood with relatively high efficiency. The purpose of this study was to compare the efficiencies of retrovirus-mediated gene transfer into early (23-28 weeks' gestation) and term (37-41 weeks' gestation) umbilical cord blood CD34(+) hematopoietic progenitor cells. STUDY DESIGN: CD34(+) cells were purified from cyropreserved early (23-28 weeks' gestation) and term (37-40 weeks' gestation) umbilical cord blood specimens with fluorescence-activated cell sorting. The CD34(+) cells were then transduced in virus-containing medium (gibbon ape leukemia virus pseudotype vector LAPSN [PG13]) in wells coated with the recombinant human fibronectin fragment CH-296 and in the presence of multiple hematopoietic growth factors (interleukin 6, stem cell factor, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, and megakaryocyte growth and development factor) and protamine sulfate. The LAPSN (PG13) virus-containing medium was changed every 12 hours for 48 hours, after which time colony-forming cells were assayed in soft agar. The gibbon ape leukemia virus pseudotype vector LAPSN (PG13) contains the human placental alkaline phosphatase and neomycin phosphotransferase (neo ) genes. The efficiency of gene transfer was assessed by histochemical staining of colony-forming cells in agar for expression of heat-stable alkaline phosphatase. RESULTS: Gene transfers, as assessed by alkaline phosphatase staining of colony-forming cells (granulocyte-macrophage colony-forming units and erythroid burst-forming units), were similar for CD34(+) hematopoietic progenitor cells from early (58.4% +/- 11.8%) and term (63.2% +/- 12.5%) gestation fetal umbilical cord blood. CONCLUSION: CD34(+) hematopoietic progenitor cells from midgestation fetal blood can be transduced with high efficiency using techniques optimized for postnatal samples with a gibbon ape leukemia virus pseudotype vector. The early fetus may be a preferable target for gene therapy because of the higher number of circulating CD34(+) and CD38(-) cells relative to term cord blood, their greater proliferative capacity, and the rapid expansion of the fetal hematopoietic system that occurs from the second trimester to delivery. Because in vitro studies of gene transfer into hematopoietic progenitor cells and long-term culture-initiation cells have not been predictive of the efficiency of gene transfer into marrow-repopulating cells in vivo, studies that examine clinically applicable approaches to in utero gene therapy in appropriate animal models are still needed.     

Differences in umbilical cord blood units collected during cesarean section,before or after the delivery of the placenta

Umbilical cord blood is largely employed as an alternative source of stem cells in the treatment of hemato-oncological diseases. Current results show that the success rate of purified umbilical cord blood engraftment is comparable to that obtained using bone marrow, and it is directly related to the number of pluripotent stem cells transplanted. The technique of fetal blood collection varies among different umbilical cord blood banks. Many authors collect umbilical cord blood during vaginal delivery, after placental detachment, while others collect it while the placenta is still within the uterus. In a previous randomized trial, we showed a greater collection of umbilical cord blood before placental detachment during vaginal delivery. The present study was performed to determine whether umbilical cord blood collection before placental detachment (group A) during cesarean section is superior to that after placental delivery (group B) puncturing the umbilical vein once and using a closed bag system. To accomplish this, 47 pregnant women subjected to cesarean section were enrolled in the study. Twenty-one of them were allocated to group A, while the remaining 26 formed group B. The volume of umbilical cord blood collected from the patients of group A was greater than that collected from patients of group B. The cord blood volume collected was 90.7 +/- 6.0 versus 60.9 +/- 13.7 ml; the cord blood nucleated cell number was 10.1 +/- 1.2 x 10(8) vs. 7.1 +/- 0.8 x 10(8); and the mean cord blood CD34+ cell number was 20.0 +/- 6.0 x 10(5) vs. 16.4 +/- 2.4 x 10(5), respectively.     

Effect of preeclampsia on umbilical cord blood hematopoietic progenitor-stem cells

OBJECTIVE: The aim of the present study was to determine the influence of preeclampsia on cord blood hematopoietic progenitor-stem cells obtained at delivery because cord blood is increasingly used clinically for stem cell retrieval as an alternative to bone marrow. STUDY DESIGN: Umbilical cord blood was collected from patients fulfilling the criteria for preeclampsia and from gestational age- and birth weight-matched control subjects at delivery (patient/control subjects ratio, 1:2). Cord blood volume and nucleated cell content were measured, and the number of hematopoietic progenitor-stem cells was determined by means of fluorescence-activated cell sorting with the CD34(+) epitope and by means of colony assays with different hematopoietic growth factors. In addition, the expression of adhesion molecules by CD34(+) progenitor-stem cells was examined. RESULTS: In pregnancies affected by preeclampsia, volume and nucleated cell and total CD34(+) cell contents in the collected cord blood were significantly smaller compared with those of control subjects. Furthermore, there was a trend toward a smaller relative number of CD34(+) cells and colony-forming units per nucleated cell in cord blood samples from preeclamptic patients. No difference in the expression of the cell-adhesion molecules leukocyte function-associated antigen 1, very late activation antigen 4, and L-selectin by CD34(+) cells could be found. CONCLUSION: This study shows that preeclampsia affects umbilical cord blood volume and nucleated cell and progenitor-stem cell numbers obtained at birth.     

In vitro cell growth stimulated by recombinant human cytokines can help to diagnose transient leukemia in neonates.

BACKGROUND/PURPOSE: In a previous study, we demonstrated that in vitro cell growth stimulated by human placental conditioned medium distinguished between transient leukemia (TL) and congenital acute myeloid leukemia (AML) in neonates. We then sought to determine whether the application can be expanded if in vitro cell growths are stimulated by recombinant human cytokines including granulocyte-macrophage colony-stimulating factor (rhGM-CSF), interleukin-3 (rhIL-3), stem cell factor (rhSCF) and thrombopoietin (rhTPO). METHODS: Eight neonates with features indistinguishable from AML were studied. Seven patients had Down syndrome and the eighth a normal phenotype. Bone marrow or peripheral blood mononuclear cells (MNC) were cultured in the presence of rhGM-CSF+rhIL-3+rhSCF or of rhTPO alone. After incubation, granulocyte-macrophage colony-forming units (CFU-GM)-derived colonies and clusters were scored on an inverted microscope. Colony-forming units-megakaryocyte (CFU-MK)-derived colonies were counted with an in situ CD61 immunostained dish. Liquid suspension cultures of MNC were stimulated by rhGM-CSF and/or rhTPO. RESULTS: CFU-GM-derived colonies and clusters from bone marrow and peripheral blood MNC revealed normal patterns in seven patients. RhTPO-stimulated megakaryocyte colony formation was normal in one patient. Cytospin smears of liquid suspension cultures all showed good myeloid or megakaryocytic maturation consistent with TL rather than AML. One neonate died on the 2nd day of life, but in the seven remaining patients, blasts disappeared from the peripheral blood within 10 months. Among four patients followed long-term, one developed myelodysplastic syndrome at 21 months. This child was given tailored chemotherapy and had a disease-free survival>20 months. CONCLUSION: In vitro cell growth stimulated by recombinant human cytokines can help to diagnose TL in neonates.     

Placing the newborn on the maternal abdomen after delivery increases the volume and CD34 cell content in the umbilical cord blood collected: an old maneuver with new applications.

OBJECTIVE: Our purpose was to increase the number of the progenitor cells in umbilical cord blood collected for transplantation. STUDY DESIGN: We randomly assessed the effect of "upper" and "lower" positions of the newborn on the volume and progenitor cell (CD34(+)) content of the umbilical cord blood collected from 49 healthy, vaginally delivered, term neonates. RESULTS: Twenty-two collections were performed in the "upper" and 27 in the "lower" position. The volume of umbilical cord blood obtained in the "upper" position was 108.1 +/- 19.1 mL compared with 42.6 +/- 19.5 mL in the "lower" position (P <.0001). Mononuclear cell separation revealed significantly higher numbers of cells in umbilical cord blood obtained in the "upper" group (P <.01). Although the percentage of CD34(+) cells was comparable, the absolute number of CD34(+) cells was significantly higher in the "upper" group because of the larger volume collected (P <.02). At 24 hours after delivery the hemoglobin levels were not significantly different between newborns of the 2 groups. CONCLUSIONS: Placing the newborn on the maternal abdomen after delivery and before cord clamping may significantly increase the volume of umbilical cord blood collected and therefore the CD34(+) counts that improve transplantation success without placing the mother or the newborn at risk.