Blood cells最新文献_第4页

Extensive proliferative capacity of single isolated CD34 human cord blood cells in suspension culture. 单个分离的CD34人脐带血细胞在悬浮培养中的广泛增殖能力。

Blood cells Pub Date : 1994-01-01

M Xiao, H E Broxmeyer, M Horie, S Grigsby, L Lu

{"title":"Extensive proliferative capacity of single isolated CD34 human cord blood cells in suspension culture.","authors":"M Xiao, H E Broxmeyer, M Horie, S Grigsby, L Lu","doi":"","DOIUrl":"","url":null,"abstract":"Nonadherent, low-density T-lymphocyte-depleted (NALT-) CD34 cells from normal human cord blood were assessed in suspension culture for the effects of recombinant cytokines on their proliferation, differentiation, and generation of myeloid progenitor cells. In this cell population, 82% of cells expressed c-kit protein as assessed by in situ hybridization, and their cloning efficiency was 85% when cells were plated at low cell numbers with combinations of growth factors. CD34 cells were sorted as 1, 5, or 10 cell(s) per well and also at 5000 cells per dish to initiate stromal-free suspension cultures in the presence of steel factor (SLF), interleukin (IL)-1 alpha, and IL-3. Forty-eight percent of the wells started with a single CD34 cell were positive for growth after 14 days, and the wells contained greater than 5 x 10(3) cells by 21-28 days. Progenitors were assayed weekly with cultures initiated with 1 or 5000 cells. While the fold expansion of nucleated cells was greater in cultures initiated with one cell per well (> 5000 compared to 791-fold expansion for 5000 cells), the fold expansion of progenitors was greater than 5000 cells were used to initiate cultures. Under optimal conditions, there was, respectively, a 160-, 164-, and 57-fold output of high proliferative potential colony-forming cells, granulocyte-macrophage colony-forming units, and erythroid burst-forming units/granulocyte erythroid macrophage megakaryocyte colony-forming units within 1-3 weeks for cultures initiated with 5000 CD34 cells compared with respective fold increases of 29, 16, and 1, for single-initiated cultures. These results demonstrate the expansion capacity of single CD34 cord blood cells and demonstrate that factors in addition to SLF, IL-1 alpha, and IL-3 are necessary for optimal expansion of progenitors from single isolated CD34 cells.","PeriodicalId":75604,"journal":{"name":"Blood cells","volume":"20 2-3","pages":"455-66; discussion 466-7"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18544575","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Comparative analysis of retroviral-mediated gene transduction into CD34+ cord blood hematopoietic progenitors in the presence and absence of growth factors. 存在和不存在生长因子时逆转录病毒介导的基因转导到CD34+脐带血造血祖细胞的比较分析。

Blood cells Pub Date : 1994-01-01

Y J Shi, R N Shen, L Lu, H E Broxmeyer

{"title":"Comparative analysis of retroviral-mediated gene transduction into CD34+ cord blood hematopoietic progenitors in the presence and absence of growth factors.","authors":"Y J Shi, R N Shen, L Lu, H E Broxmeyer","doi":"","DOIUrl":"","url":null,"abstract":"Hematopoietic stem and progenitor cells present in umbilical cord blood at the birth of a child are efficiently transduced ex vivo by genes using retroviral vectors in combination with exposure of these cells to combinations of growth factors. Because retroviral-mediated gene transduction of adult bone marrow and blood hematopoietic stem and progenitor cells is greatly enhanced by growth factors, we evaluated the possibility that cord blood progenitors, which have extensive proliferative and replating capacity, could be efficiently transduced with a TK neo gene in a retroviral vector in the absence of growth factors, and also determined the influence of exogenously added growth factors on this transduction. Highly purified CD34+ (62% pure) cord blood cells isolated by magnetic bead separation were cultured in suspension for 72 hours with viral supernatant in the absence and presence of interleukin-3 (IL-3), IL-6, and steel factor. Evaluation of progenitor cell-derived colonies and polymerase chain reaction (PCR)/Southern analysis of the TK neo gene in resultant colony cells demonstrated that some gene transduction was apparent in the absence of growth factors (12.8-14.3% by PCR), but that this was greatly enhanced (40.0-44.4%) by addition of growth factors. Reverse transcription PCR analysis of the expression of IL-3, IL-6, and granulocyte-macrophage colony-stimulating factor genes in this population of cells suggested that the transduction, although at a lower efficiency, in the absence of added growth factors might in part be due to \"constitutive\" and viral supernatant-induced expression of these cytokine genes in the CD34(+)-enriched cell population.(ABSTRACT TRUNCATED AT 250 WORDS)","PeriodicalId":75604,"journal":{"name":"Blood cells","volume":"20 2-3","pages":"517-24"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18544578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Umbilical cord blood cell transduction by retroviral vectors: preclinical studies to optimize gene transfer. 逆转录病毒载体的脐带血细胞转导:优化基因转移的临床前研究。

Blood cells Pub Date : 1994-01-01

M E Hanley, J A Nolta, R Parkman, D B Kohn

{"title":"Umbilical cord blood cell transduction by retroviral vectors: preclinical studies to optimize gene transfer.","authors":"M E Hanley, J A Nolta, R Parkman, D B Kohn","doi":"","DOIUrl":"","url":null,"abstract":"Human umbilical cord blood (UCB) can be a source of hematopoietic stem cells for gene therapy, as an alternative to allogeneic bone marrow transplantation, for the treatment of a number of genetic diseases. To determine conditions that yield maximal gene transfer into UCB progenitor cells, we examined a number of variables. We used cell-free retroviral vector supernatants that convey neomycin (G418) resistance and measured the percentage of G418-resistant progenitor-derived colonies. Adding retroviral supernatant to the UCB cells in basal medium once a day for 3 days produced a threefold increase of G418-resistant colonies (9.8%) compared to a single exposure to supernatant (3.1%). To establish whether recombinant human growth factors are beneficial during transduction, the presence of interleukin-3 (IL-3), IL-6, and mast cell growth factor (MGF, a c-kit ligand) were compared in different combinations. Inclusion of the three factors together caused a threefold increase of gene transfer (30.4%) compared to transduction in basal medium. When the UCB cells were precultured in medium containing IL-3, IL-6, and MGF for 3 days before addition of the retroviral supernatant on days 4, 5, and 6, the average extent of gene transfer was 21.8%, compared with an average of 34.4% when UCB cells were transduced on days 1, 2, and 3. The presence of marrow stroma during the transduction of the UCB cells did not further increase gene transfer. We conclude that UCB progenitor cells can be efficiently transduced with the use of recombinant human growth factors IL-3, IL-6, and MGF and may be a suitable source for gene therapy.","PeriodicalId":75604,"journal":{"name":"Blood cells","volume":"20 2-3","pages":"539-43; discussion 544-6"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18545694","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Commentary: hematopoietic progenitors in fetal and adult tissue. 评论:胎儿和成人组织中的造血祖细胞。

Blood cells Pub Date : 1994-01-01

L W Terstappen, F Lund-Johansen

引用次数: 0

Isolation and ex vivo expansion of CD34+ cells from cord blood using dextran sedimentation and avidin column selection. 右旋糖酐沉淀法和亲和素柱选择法在脐带血中分离和体外扩增CD34+细胞。

Blood cells Pub Date : 1994-01-01

S Heimfeld, D F Kalamasz, B L Fogarty, R Fei, Z N Tsui, H M Jones, R J Berenson

引用次数: 0

Proposed policies and procedures for the establishment of a cord blood bank. 建议建立脐带血银行的政策和程序。

Blood cells Pub Date : 1994-01-01

J McCullough, M E Clay, S Fautsch, H Noreen, M Segall, E Perry, D Stroncek

引用次数: 0

Commentary: cord blood banking for bone marrow reconstitution: a need for standardization and international cooperation. 评论:用于骨髓重建的脐带血库:需要标准化和国际合作。

Blood cells Pub Date : 1994-01-01

Van Rood

引用次数: 0

Engraftment of normal murine marrow into nonmyeloablated host mice. 将正常小鼠的骨髓移植到未清髓的宿主小鼠体内。

Blood cells Pub Date : 1994-01-01

P J Quesenberry, H Ramshaw, R B Crittenden, F M Stewart, S Rao, S Peters, P Becker, P Lowry, M Blomberg, J Reilly

引用次数: 0

Commentary: Immunohistochemistry represents a useful tool to study human cell engraftment in SCID mice transplantation models. 评论:免疫组织化学是研究人类细胞植入SCID小鼠移植模型的有用工具。

Blood cells Pub Date : 1994-01-01

A Orazi, S E Braun, H E Broxmeyer

引用次数: 0

Cord blood transplantation: implications for graft vs. host disease and graft vs. leukemia. 脐带血移植:移植物抗宿主病和移植物抗白血病的意义。

Blood cells Pub Date : 1994-01-01

D T Harris

{"title":"Cord blood transplantation: implications for graft vs. host disease and graft vs. leukemia.","authors":"D T Harris","doi":"","DOIUrl":"","url":null,"abstract":"Cord blood (CB) was analyzed for its alloreactive immune potential to evaluate its capacity to mediate graft vs. host disease (GVHD) and graft vs. leukemia (GVL) effects. Cord blood was observed to display minimal innate cytotoxic capacity but was capable of rapidly developing significant nonspecific natural killer cell-like effector mechanisms. Cord blood was unable to generate effective alloantigen-specific cytotoxic T lymphocytes (CTL), which was at least partially due to an altered lymphokine profile. The frequency of alloreactive T cells present in CB (whether assessed as total responding T-cell or CTL precursors) was greatly reduced as compared to adult peripheral blood lymphocytes (PBLs). However, the frequency of nonspecific effector cells was equivalent to PBLs. Significantly, CB T cells seemed to have undergone some type of developmental tolerance to maternal HLA antigens in utero, which could greatly increase the utility of CB in familial transplants. That is, CB T cells were unresponsive to noninherited maternal HLA antigens. Finally, CB demonstrated significant GVL capacity whether measured in vitro or in an animal model in vivo. Thus, the use of CB in most transplant settings should be free of the immunological problems associated with GVHD yet still be an effective mediator of GVL.","PeriodicalId":75604,"journal":{"name":"Blood cells","volume":"20 2-3","pages":"560-4; discussion 564-5"},"PeriodicalIF":0.0,"publicationDate":"1994-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"18749585","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0