Journal of hematotherapy最新文献

{"title":"Ethical issues in stem cell transplantation.","authors":"M Kletzel, E Morgan, J Frader","doi":"10.1089/scd.1.1998.7.197","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.197","url":null,"abstract":"","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 3","pages":"197-203"},"PeriodicalIF":0.0,"publicationDate":"1998-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.197","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20540534","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}

{"title":"Increased transduction efficiency of primary hematopoietic cells by physical colocalization of retrovirus and target cells.","authors":"M Hutchings,&nbsp;K Moriwaki,&nbsp;D Dilloo,&nbsp;T Hoffmann,&nbsp;S Kimbrough,&nbsp;H E Johnsen,&nbsp;M K Brenner,&nbsp;H E Heslop","doi":"10.1089/scd.1.1998.7.217","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.217","url":null,"abstract":"<p><p>Efficient gene transfer into hematopoietic stem cells offers a number of potential therapeutic applications. However, the relatively low titer of retroviral supernatants and the requirement for cell division to ensure integration have meant that transduction efficiency has been low. We have modified a flowthrough approach to cell transduction and have been able consistently to increase gene transfer efficiency into human hematopoietic progenitor cells. We transduced CD34 cells with retroviral vectors encoding a truncated nerve growth factor receptor (NGFR) or neo. Retroviral supernatant was pulled through 0.2-micron polycarbonated membranes, followed by placement of cells on the filter. In the absence of cytokines, the transduction efficiency of CD34 cells with a NGFR vector was increased 3-11-fold over that obtained at an identical MOI in liquid culture to produce 11%-44% transduction. Furthermore, both Thy1+ and Thy1- subsets in a total CD34 population were transduced with similar efficiency, and transduction with a neo vector, as measured by G418 resistance in clonogenic assays, increased 1.5-5-fold. The mechanism by which gene transfer is improved may reflect colocalization of cells and retrovirus. Costaining of cells transduced on the filter with an NGFR retrovirus with both an NGFR antibody and a gp70 antibody that recognizes viral coat protein revealed high-level coexpression. The levels of in vitro gene transfer we obtain are equivalent to those observed when CD34 cells are cocultured in liquid culture with cytokines. However, culture with cytokines may commit CD34 cells to differentiation and has produced disappointingly low levels of subsequent in vivo gene transfer. Gene marking studies using distinguishable retroviral vectors will provide a means of learning whether the effects of flowthrough transduction genuinely enhance the efficiency of gene transfer to human marrow-repopulating cells.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 3","pages":"217-24"},"PeriodicalIF":0.0,"publicationDate":"1998-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.217","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20540536","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}

{"title":"Second attempts at mobilization of peripheral blood stem cells in patients with initial low CD34+ cell yields.","authors":"C H Weaver,&nbsp;K Tauer,&nbsp;B Zhen,&nbsp;L S Schwartzberg,&nbsp;B Hazelton,&nbsp;Z Weaver,&nbsp;L Lewkow,&nbsp;C Allen,&nbsp;K Longin,&nbsp;C D Buckner","doi":"10.1089/scd.1.1998.7.241","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.241","url":null,"abstract":"<p><p>The purpose of this study was to determine the effectiveness of second mobilization strategies in patients who yielded < 2.5 x 10(6) CD34+ PBSC/kg after initial mobilization. Repeat mobilization attempts were made with chemotherapy and G-CSF (n = 61) or G-CSF alone (n = 58) in patients who failed initial mobilization with chemotherapy and G-CSF (n = 92) or G-CSF alone (n = 27). A median of 0.27 x 10(6) CD34+ cells/kg per apheresis was collected after the second mobilization, compared with 0.16 with initial harvests (p = 0.0001). Forty-eight percent achieved a target CD34+ cell dose > or = 2.5 x 10(6)/kg when harvests from the first and second mobilizations were combined. Fifteen of 17 patients (88%) with > or = 1.5 x 10(6) CD34+ cells/kg harvested after first mobilization had > or = 2.5 x 10(6) CD34+ cells/kg collected when first and second harvests were combined, as compared with 42 of 102 (41%) achieving < 1.5 x 10(6) CD34+ cells/kg with first PBSC harvests (p = 0.0001). Second mobilizations with chemotherapy and G-CSF or G-CSF alone resulted in similar CD34+ cell yields. Toxicities of second mobilizations were comparable with those of first mobilizations. Seventy-nine patients (66%) received high-dose chemotherapy with PBSC support, with recovery of neutrophils and platelets in a median of 11 and 15 days, respectively. Transplant-related mortality was 4%, and event-free survival at 2 years was 0.34. It was concluded that second mobilization attempts in patients who fail to achieve > or = 2.5 x 10(6) CD34+ cells/kg on initial mobilization were successful in 48% of patients. G-CSF alone was as effective as chemotherapy plus G-CSF in mobilizing CD34+ cells and was associated with less morbidity.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 3","pages":"241-9"},"PeriodicalIF":0.0,"publicationDate":"1998-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.241","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20540538","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}

{"title":"Distribution of mobilized progenitor cells in the buffy coat of the haemonetics MCS3p cell separator: a study to optimize the collection of progenitors by leukapheresis.","authors":"M Ghielmini,&nbsp;U Pfister,&nbsp;E Zucca,&nbsp;S van den Bosch,&nbsp;P Tamásy,&nbsp;G Marangoni,&nbsp;G Bertoli,&nbsp;J L Derivaz,&nbsp;F Cavalli","doi":"10.1089/scd.1.1998.7.251","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.251","url":null,"abstract":"<p><p>Hematopoietic progenitor and stem cells for transplantation can be mobilized into the circulation and collected by leukapheresis. In this procedure, the leukocytes are distributed in the buffy coat along a density gradient, and the composition of the final product depends on which layer was collected. For the Haemonetics MCS3p Cell Separator, the manufacturer recommended starting the progenitor cell collection at a light transmission of 30%-40% (compared with plasma) and continue it for 40-50 ml. To optimize the use of this machine, the buffy coat it produces was studied in 12 patients by collecting it in fractions of increasing specific weight. Each fraction was analyzed by morphology, immunocytometry, and cell culture. We found that the buffy coat uniformly contains 8 times more leukocytes than blood, but the proportion of each white cell type varies along a gradient. The lymphocyte-predominant lighter layers are richer in CD34+ cells when compared with the granulocyte-predominant denser layers (6-14 times versus 2-4 times more than blood). The majority of CD34+ cells are found at a light transmission of 10%-70% (hematocrit 6-9). We conclude that cells for transplantation should be collected in a lighter fraction of the buffy coat than originally suggested by the manufacturer.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 3","pages":"251-6"},"PeriodicalIF":0.0,"publicationDate":"1998-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.251","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20540539","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}

{"title":"A closed culture system for the ex vivo transduction and expansion of human T lymphocytes.","authors":"E Robinet,&nbsp;J M Certoux,&nbsp;C Ferrand,&nbsp;P Maples,&nbsp;A Hardwick,&nbsp;J Y Cahn,&nbsp;C W Reynolds,&nbsp;W Jacob,&nbsp;P Hervé,&nbsp;P Tiberghien","doi":"10.1089/scd.1.1998.7.205","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.205","url":null,"abstract":"<p><p>A phase I clinical trial is currently being performed at our institution, with the aim of evaluating the feasibility and toxicity related to the administration of herpes simplex thymidine kinase gene-expressing human primary T lymphocytes following allogeneic hematopoietic stem cell transplantation. The need for safe and standardized preparation conditions for gene-modified cells is crucial. We describe the closed culture system used in the current trial for ex vivo retroviral-mediated gene transfer and transduced cell selection. Cell handling is performed in closed systems using a sterile connection device that avoids opening the culture system. Cell numbers during the production process increased from 93 +/- 16 on day 0 to 440 +/- 92 x 10(6) on day 12 (7.2 +/- 1.4-fold increase) (n = 11). Transduction efficiency before and after G418 resistance-based selection was 13.5 +/- 3.8% and 90.0 +/- 1.4%, respectively. Safety and efficacy testing included a search for replication-competent retrovirus, endotoxins, Mycoplasma, and bacterial contamination (n = 0/9), PCR-DNA, % CD3+ cells (91 +/- 2%), and viability after thawing (82 +/- 3%). Effective working time from day 0 to day 12 is approximately 20 h. The closed system we developed allows for safe and reproducible ex vivo preparation of gene-modified primary T lymphocytes for clinical use.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 3","pages":"205-15"},"PeriodicalIF":0.0,"publicationDate":"1998-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.205","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20540535","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}

{"title":"Retroviral-mediated marker gene transfer in hematopoiesis-supportive marrow stromal cells.","authors":"C E Bulabois,&nbsp;V Yerly-Motta,&nbsp;B T Mortensen,&nbsp;P Fixe,&nbsp;J P Remy-Martin,&nbsp;P Hervé,&nbsp;P Tiberghien,&nbsp;P Charbord","doi":"10.1089/scd.1.1998.7.225","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.225","url":null,"abstract":"<p><p>A Moloney-derived retrovirus containing both LacZ and NeoR genes (G1BgSVNa from Genetic Therapy, Inc.), was used to transduce human and murine bone marrow stromal cells. Different kinds of stromal cells that were able to support hematopoiesis were transduced by incubation for 24 h in the presence of virus-containing supernatant. Semiconfluent layers of MRC-5 (human, myofibroblastic, fetal, pulmonary) and MS-5 (murine, myofibroblastic, medullary) cells were successfully transduced after one 24-h incubation, as demonstrated by G418 resistance and Escherichia coli beta-galactosidase staining. In contrast, human stromal cells, purified from primary confluent layers grown for 3-4 weeks, could not be transduced. However, stromal cells generated after 10-12 days in culture from Stro-1+ and 1B10+ stromal precursors were successfully transduced in the presence of basic fibroblast growth factor. Transduced stromal cells maintained a myofibroblastic phenotype, although with a decreased number of alpha-SM actin-positive microfilaments in MS-5 cells. The ability to support the generation of stroma-adherent colony-forming cells from cocultured cord blood CD34+ cells after 4 weeks in culture was similar before and after transduction and G418 selection. In conclusion, human primary stromal precursors can be efficiently transduced, and the stromal cell phenotype and function are not significantly altered after retroviral-mediated transfer of marker genes.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 3","pages":"225-39"},"PeriodicalIF":0.0,"publicationDate":"1998-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.225","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20540537","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}

{"title":"Long-term cryopreservation: successful trilineage engraftment after autologous bone marrow transplantation with bone marrow cryopreserved for seven years.","authors":"A Re,&nbsp;K Vijayaraghavan,&nbsp;M M Basade,&nbsp;S He,&nbsp;S C Gulati","doi":"10.1089/scd.1.1998.7.185","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.185","url":null,"abstract":"<p><p>Successful autologous bone marrow transplantation (ABMT) and peripheral blood stem cell transplantation depend on safe hematopoietic stem cell (HSC) cryopreservation and storage. Several successful methods for cryopreservation and storage have been established and are commonly used all over the world. However, little is known about the effects of long-term cryopreservation on the capacity to sustain a complete immunohematopoietic engraftment. Several authors have investigated stem cell viability after cryopreservation and storage for more than 5 years and reported preclinical good viabilities in terms of dye-exclusion or colony-forming capability in vitro. Only two studies using BM cryopreserved for more than 5 years for transplantation are reported, but they did not provide proof of trilineage engraftment. In February 1997 at our institution, a patient with relapsed non-Hodgkin's lymphoma underwent ABMT with BM harvested in February 1990. He achieved a granulocyte count > 500 x 10(6)/L on day 21 and a self-supporting platelet count > 20 x 10(9)/L on day 30. After day 29, his hemoglobin level was satisfactory without need of transfusion support. This successful trilineage engraftment with cryopreserved BM that had been stored for 7 years suggests that HSC are able to maintain their capability to reconstitute hematopoiesis for a long time.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 2","pages":"185-8"},"PeriodicalIF":0.0,"publicationDate":"1998-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.185","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20518014","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}

{"title":"Not all healthy donors mobilize hematopoietic progenitor cells sufficiently after G-CSF administration to allow for subsequent CD34 purification of the leukapheresis product.","authors":"M Holm","doi":"10.1089/scd.1.1998.7.111","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.111","url":null,"abstract":"","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 2","pages":"111-3"},"PeriodicalIF":0.0,"publicationDate":"1998-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.111","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20518105","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}

{"title":"Effect of stromal cell coculture on progenitor cell expansion and myeloid effector function in vitro.","authors":"J L Liesveld,&nbsp;B A Martin,&nbsp;A W Harbol,&nbsp;K E Rosell,&nbsp;T J Belanger,&nbsp;S E Reykdal,&nbsp;D L Plekavich,&nbsp;C N Abboud","doi":"10.1089/scd.1.1998.7.127","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.127","url":null,"abstract":"<p><p>Stimulation of CD34(+)-enriched marrow or light density marrow with various growth factor combinations can generate granulocyte progenitors and mature neutrophils in vitro. In this work, we have examined the influence of irradiated marrow stromal layers on growth factor-induced myeloid and early multipotential progenitor expansion from enriched marrow CD34+ progenitors. We have also explored whether the addition of early-acting growth factors known to enhance myelopoiesis in long-term culture, such as fibroblast growth factor (b-FGF), insulin growth factor (IGF-1), c-kit ligand or stem cell factor (SCF), and flk-2flt-3 ligand (FL), can lengthen survival of CD34+ progenitors in these cultures. Stromal cell coculture resulted in greater numbers of total cells and CFU-GM at day 7 and day 14, but with the addition of multiple growth factors, these effects of stromal cell coculture were diminished. At day 14, generally < 1% of the expanded cells over stromal coculture conditions were CD34+, with up to 90% demonstrating CD15 positivity. Culture of CD34+ cells in the presence of early-acting growth factors did not cause significant expansion of CD34+ cells over a 14-day life span, even in the presence of marrow stromal cells. These data suggest that although stromal cell coculture for a period up to 14 days can enhance expansion of total cell numbers and CFU-GMs, stromal cell presence does not lead to expansion of CD34+ cells in these cultures and may diminish the number of clonogenic cells present when growth factors with differentiating capacity are present. Mature neutrophils harvested from such cultures are capable of chemotaxis, actin polymerization, and migration, suggesting a replete functional status.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 2","pages":"127-39"},"PeriodicalIF":0.0,"publicationDate":"1998-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.127","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20518008","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}

{"title":"CD19 selection improves the sensitivity of B cell lymphoma detection.","authors":"R E Pugh,&nbsp;M A Bitter,&nbsp;E J Shpall,&nbsp;L S Hami,&nbsp;D M Wolf,&nbsp;W A Franklin","doi":"10.1089/scd.1.1998.7.159","DOIUrl":"https://doi.org/10.1089/scd.1.1998.7.159","url":null,"abstract":"<p><p>Reinfusion of residual tumor cells into B cell non-Hodgkin's lymphoma (B-NHL) patients during autologous transplantation may be an important cause of disease relapse. Determining the extent to which B-NHL cells are present in autologous progenitor cell products and if the presence of residual B-NHL cells is predictive of relapse will require extremely sensitive methods of detecting rare B-NHL cells. We attempted to improve the sensitivity of polymerase chain reaction (PCR)-based detection of rare B-NHL cells by preselecting CD19+ cells using an immunomagnetic column. To measure detection sensitivity, we prepared samples containing different levels of B-NHL cell contamination by mixing B-NHL cell lines containing the chromosomal translocation t(14;18) bcl-2/JH) with control leukapheresis samples. DNA extracted from each CD19-selected sample and from each matched nonselected sample was added to a PCR to amplify the bcl-2/JH breakdown junction. CD19 preselection improved the sensitivity of detection of t(14;18)-positive B-NHL cells 115-fold, so that B-NHL cells at a concentration of 1 tumor cell per 1 x 10(6) hematopoietic cells were detected in every specimen evaluated. t(14;18)-positive cells were not detected in any of 13 control leukapheresis specimens. We conclude that a combination of CD19 preselection and PCR amplification may improve the sensitivity of detection of rare lymphoma cells by two orders of magnitude without a significant decrease in specificity.</p>","PeriodicalId":77208,"journal":{"name":"Journal of hematotherapy","volume":"7 2","pages":"159-68"},"PeriodicalIF":0.0,"publicationDate":"1998-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/scd.1.1998.7.159","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"20518011","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}