Cobblestone Area-Forming Cell (CAFC) Content of Cord Blood Does Not Predict Engraftment in Double Cord Blood Transplantation.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2243-2243
Author(s):  
Simon N. Robinson ◽  
Marcos de Lima ◽  
Hong Yang ◽  
William K. Decker ◽  
Dongxia Xing ◽  
...  
Keyword(s):  
Cord Blood ◽  
Ex Vivo ◽  
Cord Blood Transplantation ◽  
Randomized Study ◽  
Clinical Samples ◽  
Hematopoietic Stem ◽  
Blood Transplantation ◽  
Qualitative Measure

Abstract INTRODUCTION: Delayed engraftment remains a serious problem following cord blood (CB) transplantation. It may be due, at least in part, to the limited dose of CB hematopoietic stem and progenitor cells (HSPC) transplanted. Limitations associated with HSPC dose may be reduced by the transplantation of 2 CB units. In an accompanying abstract, de Lima et al. report in detail on an ongoing MDACC randomized study in which patients received double CB units as either 2 unmanipulated units (2×UN), or 1 unmanipulated and 1 ex vivo expanded CB unit (UN+EX). This study has revealed that one CB unit ultimately predominates as the source of long-term, sustained hematopoiesis. We hypothesized that the number of primitive HSPC in the CB unit might ultimately predict which CB unit would ultimately prevail. The in vitro cobblestone area-forming cell (CAFC) assay was used to provide a measure of primitive components of the CB HSPC in each unit. A photomicrograph of a typical cobblestone area (CA) derived from a single CB-derived CAFC is shown. (Figure) CA persisting in in vitro culture for ≥6 weeks (derived from CAFCwk6) represent relatively primitive HSPC and their numbers may provide a qualitative profile for CB units. We hypothesized that of the two CB units transplanted, the one with the greater number of CAFCwk6 would ultimately persist long-term in the patient. METHODS: Clinical samples of 2×UN or UN+EX CB cells were plated in the in vitro CAFC assay.1 The frequency of CAFCwk6 was estimated and total CAFCwk6 numbers transplanted for each CB unit calculated. CAFCwk6 data for each CB unit and engraftment data from patients were analyzed to determine whether the number of CAFCwk6 transplanted was predictive of which CB unit would ultimately be responsible for long-term, sustained engraftment. RESULTS: Preliminary data was accrued from 10 patients.(Table) Six patients received 2×UN and 4 patients received UN+EX (EX indicated by #). CAFCwk6 content at transplant was predictive of which CB unit would ultimately be responsible for long-term, sustained engraftment in only 6 of the 10 cases (60%). CONCLUSION: CAFCwk6 represent a relatively primitive component of the HSPC compartment, however, these data suggest that CAFCwk6 numbers, although possibly a qualitative measure, are not a predictor of long-term sustained engraftment. Figure Figure

scholarly journals A Murine Model for Human Cord Blood Transplantation: Near-Term Fetal and Neonatal Peripheral Blood Cells Can Achieve Long-Term Bone Marrow Engraftment in Sublethally Irradiated Adult Recipients

Blood ◽  
1997 ◽  
Vol 89 (3) ◽  
pp. 1089-1099 ◽  
Author(s):  
Andromachi Scaradavou ◽  
Luis Isola ◽  
Pablo Rubinstein ◽  
Yelena Galperin ◽  
Vesna Najfeld ◽  
...  
Keyword(s):  
Cord Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Cultured Cells ◽  
Ex Vivo ◽  
Cord Blood Transplantation ◽  
Blood Transplantation ◽  
Near Term ◽  
Ex Vivo Culture

Abstract The purposes of the research reported here were first to explore a murine model for human placental and umbilical cord blood transplantation and second to evaluate the engraftment ability of ex vivo cultured hematopoietic cells. Murine near-term fetal and neonatal peripheral blood (FNPB) cells, genetically marked with the human multiple drug resistance transgene (MDR1) were used for syngeneic transplants into sublethally irradiated adult mice. Donor cells were transplanted either fresh and untreated, or after ex vivo culture in the presence of the hematopoietic growth factors recombinant murine stem cell factor, recombinant human interleukin-3 (rHu IL-3), and rHu IL-6, in a liquid culture system. To evaluate, count, and characterize FNPB progenitor cell-derived colonies, neonatal mouse mononuclear cells were cultured directly in methylcellulose with growth factors. To assess their ex vivo expansion ability, FNPB mononuclear cells were first cultured in liquid medium for 3 to 8 days and then transferred to semisolid assay plates. Evaluation of the cell counts after liquid culture showed a 1.4- to 11.6-fold increase, and the numbers of colonies observed in methylcellulose were similar to those produced by fresh FNPB cells. Donor-type engraftment was demonstrated by polymerase chain reaction (PCR) amplification of the human MDR1 transgene in the peripheral blood of all surviving animals (5 of 7 recipients of the fresh, and 3 of 8 recipients of the ex vivo–cultured cells) 2 to 4 months after transplantation. The proportion of donor leukocytes in the peripheral blood of the recipients (chimerism) was evaluated using fluorescence in situ hybridization (FISH) analysis 4 to 6 months after transplantation and ranged from 2% to 26%. In addition, bone marrow cultures were obtained from two recipient animals: one had received fresh-untreated cells and was evaluated 8 months after transplant, the other had received ex vivo cultured cells and was tested 14 months after grafting. The derived hematopoietic colonies were tested by PCR and the transgene was detected, conclusively proving long-term engraftment of donor cells. These results indicate that FNPB transplants can be successfully performed in sublethally irradiated mice with and without ex vivo culture. Long-term donor-type engraftment with sustained chimerism has been demonstrated. Thus, murine neonatal blood grafts can be used as an animal model for cord blood transplantation for gene therapy studies where complete myeloablation is not desirable and partial replacement of defective marrow may be sufficient. Furthermore, the possibility of numerically expanding hematopoietic progenitor cells contained in neonatal blood without affecting their engraftment ability could facilitate use of cord blood grafts in adult recipients.

scholarly journals Risk Factors and Options to Improve Engraftment in Unrelated Cord Blood Transplantation

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Anna D. Petropoulou ◽  
Vanderson Rocha
Keyword(s):  
Cord Blood ◽  
Hematopoietic Cell Transplantation ◽  
Ex Vivo ◽  
Cord Blood Transplantation ◽  
Clinical Problem ◽  
Alternative Source ◽  
Hematopoietic Stem ◽  
Blood Transplantation ◽  
Donor Choice ◽  
Unrelated Cord Blood

Use of umbilical unrelated cord-blood (UCB) cells as an alternative source of hematopoietic cell transplantation has been widely used mainly for patients lacking an HLA-matched donor. UCB present many advantages over bone marrow or mobilized peripheral blood from volunteer donors, such as rapid availability, absence of risk for the donor, and decreased incidence of acute graft-versus-host disease. However, a significant clinical problem is delayed engraftment that is directly correlated with the number of hematopoietic stem cells in a cord-blood unit. The identification of prognostic factors associated with engraftment that can be easily modified (e.g., strategies for donor choice) and the development of new approaches including use of multiple donors, intrabone injection of UCB, ex vivo expansion, and cotransplantation with accessory cells are of crucial importance in order to circumvent the problem of delayed engraftment after UCB transplantation. Those approaches may increase the quality and availability of UCB for transplantation.

Cytokine-Induced Killer Cells Facilitate Immune Reconstitution After Allogeneic BMT In Mice.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3719-3719
Author(s):  
Shintaro Mase ◽  
Ryosei Nishimura ◽  
Rie Kuroda ◽  
Hideaki Maeba ◽  
Kazuhito Naka ◽  
...  
Keyword(s):  
Cord Blood ◽  
Graft Failure ◽  
Immune Reconstitution ◽  
Conflicts Of Interest ◽  
Ex Vivo ◽  
Cord Blood Transplantation ◽  
Early Time ◽  
Hematopoietic Stem ◽  
Cik Cells ◽  
Blood Transplantation

Abstract Abstract 3719 Cytokine-induced killer (CIK) cells are ex vivo–expanded T lymphocytes expressing both natural killer (NK)– and T-cell markers. We have reported that adoptive transfer of allogeneic CIK cells in a murine model caused minimal graft versus host disease (GVHD) with retention of antitumor activity mediated by NKG2D, which is an activating receptor expressed on NK cells. With the purpose of potential application of CIK cells in a clinical hematopoietic stem cell transplantation, the problem we have to consider next is whether CIK cells could promote an engraftment and facilitate an immune reconstitution. To this end, lethally irradiated BALB/c mice were injected with minimal number of MHC incompatible C57BL/6 bone marrow (BM) cells alone, in which almost half of mice died because of graft failure, or with CIK concurrently. The mice receiving BM plus CIK cells survived with 93% without GVHD, demonstrating that CIK cells significantly promote an engraftment (P<0.05). In particular, recovery of CD3+ T-cells was significantly faster (p<0.05) in the mice receiving BM plus CIK cells than those receiving BM cells alone. Next we further evaluated whether CIK cells also promote an engraftment in the mice receiving non-myeloablative conditioning using low dose total body irradiation. As expected, CIK cells promoted an engraftment and favored immunoreconstitution, especially from the early time point after BMT. In conclusion, our study clearly demonstrated that CIK cells promote an engraftment and facilitate an immure reconstitution without GVHD. As recent studies demonstrated that sufficient number of CIK cells could be expanded even from washouts of cord blood units bags, infusion of CIK cells would be a potent strategy for preventing a graft-failure in clinical settings, especially after cord blood transplantation. Disclosures: No relevant conflicts of interest to declare.

scholarly journals The BET inhibitor CPI203 promotes ex vivo expansion of cord blood long-term repopulating HSCs and megakaryocytes

Blood ◽  
2020 ◽  
Vol 136 (21) ◽  
pp. 2410-2415 ◽  
Author(s):  
Peng Hua ◽  
Joanna Hester ◽  
George Adigbli ◽  
Rong Li ◽  
Bethan Psaila ◽  
...  
Keyword(s):  
Cord Blood ◽  
Progenitor Cells ◽  
Ex Vivo ◽  
Ex Vivo Expansion ◽  
Human Cord Blood ◽  
Hematopoietic Stem ◽  
Bet Inhibitor ◽  
Functional Studies

Abstract Although cytokine-mediated expansion of human hematopoietic stem cells (HSCs) can result in high yields of hematopoietic progenitor cells, this generally occurs at the expense of reduced bone marrow HSC repopulating ability, thereby limiting potential therapeutic applications. Because bromodomain-containing proteins (BCPs) have been demonstrated to regulate mouse HSC self-renewal and stemness, we screened small molecules targeting various BCPs as potential agents for ex vivo expansion of human HSCs. Of 10 compounds tested, only the bromodomain and extra-terminal motif inhibitor CPI203 enhanced the expansion of human cord blood HSCs without losing cell viability in vitro. The expanded cells also demonstrated improved engraftment and repopulation in serial transplantation assays. Transcriptomic and functional studies showed that the expansion of long-term repopulating HSCs was accompanied by synchronized expansion and maturation of megakaryocytes consistent with CPI203-mediated reprogramming of cord blood hematopoietic stem and progenitor cells. This approach may therefore prove beneficial for ex vivo gene editing, for enhanced platelet production, and for the improved usage of cord blood for transplantation research and therapy.

scholarly journals Cord Blood-Derived Hematopoietic Stem/Progenitor Cells: Current Challenges in Engraftment, Infection, andEx VivoExpansion

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Katsuhiro Kita ◽  
Jong O. Lee ◽  
Celeste C. Finnerty ◽  
David N. Herndon
Keyword(s):  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Ex Vivo ◽  
Cord Blood Transplantation ◽  
Short Review ◽  
Technical Improvement ◽  
Hematopoietic Stem ◽  
Blood Transplantation ◽  
Unrelated Donors

Umbilical cord blood has served as an alternative to bone marrow for hematopoietic transplantation since the late 1980s. Numerous clinical studies have proven the efficacy of umbilical cord blood. Moreover, the possible immaturity of cells in umbilical cord blood gives more options to recipients with HLA mismatch and allows for the use of umbilical cord blood from unrelated donors. However, morbidity and mortality rates associated with hematopoietic malignancies still remain relatively high, even after cord blood transplantation. Infections and relapse are the major causes of death after cord blood transplantation in patients with hematopoietic diseases. Recently, new strategies have been introduced to improve these major problems. Establishing better protocols for simple isolation of primitive cells andex vivoexpansion will also be very important. In this short review, we discuss several recent promising findings related to the technical improvement of cord blood transplantation.

CD3 Antibody Treatment Prevents Gvhd and Allows For Rapid, Long Term, Multilineage Engraftment Of Unfractionated UCB Into Immunodeficient Mice

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 2018-2018
Author(s):  
Mark Wunderlich ◽  
Ryan Brooks ◽  
Garrett Rhyasen ◽  
James C. Mulloy
Keyword(s):  
Stem Cell ◽  
T Cell ◽  
Cord Blood ◽  
Cord Blood Transplantation ◽  
Antibody Treatment ◽  
Hematopoietic Stem ◽  
Nsg Mice

Abstract NSG mice are currently the best hosts for human hematopoietic stem cell xenografts, due in large part to their ability to initiate robust grafts comprised of diverse human blood cell lineages. Unfortunately, NSG mice are susceptible to GVHD induced by contaminating T cells. Therefore, stem and progenitor cells must be selected with a high degree of purity. This not only complicates the procedure, but also prevents modeling of whole umbilical cord blood (WUCB) and bone marrow transplantation. Here, we have explored both in vivo and in vitro methods to prevent xenogenic GVHD utilizing the anti-T cell antibodies ATG and OKT3. IP injection of either antibody within the first 72 hours of transplant proved effective at preventing T cell expansion. Alternatively, a simple short-term incubation of the cell suspension with antibody was equally effective. ATG, but not OKT3, had significant non-specific activity resulting in reduction and even prevention of the non-T cell portion of the graft, including SCID repopulating cells, using either the in vivo or in vitro approach. However, using the OKT3 in vitro incubation protocol, we were able to initiate stable, long term, multi-lineage engraftment from whole, unselected UCB without GVHD. Additionally, WUCB transplanted mice engrafted more readily, with a wider array of human cell types in the peripheral blood as early as 2 weeks, relative to CD34+ engrafted mice. Stem cell engraftment was confirmed by successful transplant of the human graft to secondary recipients. Limiting dilution experiments showed that significant human grafts could be generated with as few as 1 million total WBCs per mouse, indicating that very large cohorts of mice can be generated with a single UCB sample. This protocol not only has the potential to significantly streamline hematopoietic xenograft studies while greatly reducing the cost and time commitments involved, but also to allow for whole cord blood transplantation modeling and generation of unique subsets of human cells in vivo. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Extensive Amplification and Self-Renewal of Human Primitive Hematopoietic Stem Cells From Cord Blood

Blood ◽  
1997 ◽  
Vol 89 (8) ◽  
pp. 2644-2653 ◽  
Author(s):  
Wanda Piacibello ◽  
Fiorella Sanavio ◽  
Lucia Garetto ◽  
Antonella Severino ◽  
Daniela Bergandi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Cord Blood ◽  
Ex Vivo ◽  
Attractive Alternative ◽  
Hematopoietic Tissue ◽  
Hematopoietic Growth Factors ◽  
Hematopoietic Stem

Abstract The use of umbilical cord blood as a source of marrow repopulating cells for the treatment of pediatric malignancies has been established. Given the general availability, the ease of procurement, and progenitor content, cord blood is an attractive alternative to bone marrow or growth factor mobilized peripheral blood cells as a source of transplantable hematopoietic tissue. However, there is a major potential limitation to the widespread use of cord blood as a source of hematopoietic stem cells for marrow replacement and gene therapy. There may be enough hematopoietic stem cells to reconstitute children, but the ability to engraft an adult might require ex vivo manipulations. We describe an in vitro system in which the growth of cord blood CD34+ cells is sustained and greatly expanded for more than 6 months by the simple combination of two hematopoietic growth factors. Progenitors and cells belonging to all hematopoietic lineages are continuously and increasingly generated (the number of colony-forming unit–granulocyte-macrophage [CFU-GM] present at the end of 6 months of culture are well over 2,000,000-fold the CFU-GM present at the beginning of the culture). Very primitive hematopoietic progenitors, including long-term culture-initiating cells (LTC-ICs) and blast cell colony-forming units, are also greatly expanded (after 20 weeks of liquid culture, LTC-IC number is over 200,000-fold the initial number). The extremely prolonged maintenance and the massive expansion of these progenitors, which share many similarities with murine long-term repopulating cells, suggest that extensive renewal and little differentiation take place. This system might prove useful in diverse clinical settings involving treatment of grown-up children and adults with transplantation of normal or genetically manipulated hematopoietic stem cells.

scholarly journals The Role of HLA in Cord Blood Transplantation

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Catherine Stavropoulos-Giokas ◽  
Amalia Dinou ◽  
Andreas Papassavas
Keyword(s):  
Cord Blood ◽  
Cord Blood Transplantation ◽  
Term Survival ◽  
Hematopoietic Stem ◽  
Graft Versus Host ◽  
Blood Transplantation ◽  
Survival Studies ◽  
Hla Mismatches

In recent years, umbilical cord blood (CB), a rich source of hematopoietic stem cells (HSC), has been used successfully as an alternative HSC source to treat a variety of hematologic, immunologic, genetic, and oncologic disorders. CB has several advantages, including prompt availability of the transplant, decrease of graft versus host disease (GVHD) and better long-term immune recovery, resulting in a similar long-term survival. Studies have shown that some degree of HLA mismatches is acceptable. This review is intended to outline the main aspects of HLA matching in different settings (related, pediatric, adult, or double-unit HSCT), its effect on transplantation outcome and the role of HLA in donor selection.

Strategies to Protect Hematopoietic Stem Cells from Culture-Induced Stress Conditions

2020 ◽  
Vol 15 ◽  
Author(s):  
Fatima Aerts-Kaya
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Ex Vivo ◽  
Stress Conditions ◽  
Stress Factors ◽  
Hematopoietic Stem ◽  
Induced Stress

: In contrast to their almost unlimited potential for expansion in vivo and despite years of dedicated research and optimization of expansion protocols, the expansion of Hematopoietic Stem Cells (HSCs) in vitro remains remarkably limited. Increased understanding of the mechanisms that are involved in maintenance, expansion and differentiation of HSCs will enable the development of better protocols for expansion of HSCs. This will allow procurement of HSCs with long-term engraftment potential and a better understanding of the effects of the external influences in and on the hematopoietic niche that may affect HSC function. During collection and culture of HSCs, the cells are exposed to suboptimal conditions that may induce different levels of stress and ultimately affect their self-renewal, differentiation and long-term engraftment potential. Some of these stress factors include normoxia, oxidative stress, extra-physiologic oxygen shock/stress (EPHOSS), endoplasmic reticulum (ER) stress, replicative stress, and stress related to DNA damage. Coping with these stress factors may help reduce the negative effects of cell culture on HSC potential, provide a better understanding of the true impact of certain treatments in the absence of confounding stress factors. This may facilitate the development of better ex vivo expansion protocols of HSCs with long-term engraftment potential without induction of stem cell exhaustion by cellular senescence or loss of cell viability. This review summarizes some of available strategies that may be used to protect HSCs from culture-induced stress conditions.

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