scholarly journals Rapid Determination of Potency in Cord Blood and in Mobilized Peripheral Blood Stem Cells by Flow Cytometry

2019 ◽  
Vol 8 (S1) ◽  
Author(s):  
Carl Simard ◽  
Sonia Néron ◽  
Patrick Trépanier ◽  
Nellie Dumont ◽  
Diane Fournier
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Cord Blood ◽  
Peripheral Blood ◽  
Rapid Determination ◽  
Peripheral Blood Stem Cells ◽  
Blood Stem Cells ◽  
Mobilized Peripheral Blood

Granulocyte-Derived Cationic Peptides (GDCPs) Present in Leucophoresis Products Enhance Homing of Hematopoietic Stem Cells (HSCs) to SDF-1 Gradient; Potential Implications for Accelerated Recovery of Hematopoiesis After Transplantation of Mobilized Peripheral Blood Stem Cells (PBSC).

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 371-371
Author(s):  
HakMo Lee ◽  
Wan Wu ◽  
Marcin Wysoczynski ◽  
Magdalena Kucia ◽  
Mary J. Laughlin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Peripheral Blood ◽  
Ex Vivo ◽  
Cationic Peptides ◽  
Peripheral Blood Stem Cells ◽  
Hematopoietic Stem ◽  
Blood Stem Cells ◽  
Anaphylatoxin C3a ◽  
Mobilized Peripheral Blood

Abstract Abstract 371 Current strategies to accelerate hematopoietic reconstitution after transplantation, include transplantation of greater numbers of HSC or ex vivo expansion of harvested HSC before transplant. However, the number of HSC availabel for allogeneic or autologous transplantation can be low (e.g., umbilical cord blood, poor mobilizers) and strategies to expand HSC and maintain equivalent engraftment capability ex vivo are limited. We reported that some compounds present in leucopheresis products [(e.g., platelet-derived microparticles (Blood 2001, 98: 3143)] and some complement cascade cleavage fragments, e.g., anaphylatoxin C3a (Blood 2005, 101, 3784), enhance the homing responses of HSC to SDF-1 gradient. We recently noted that small cationic peptides released from activated granulocytes (beta2-defensin and cathelicidin) positively prime responsiveness of murine and human HSC to SDF-1 gradient (Leukemia 2009; in press). Accordingly, both compounds enhanced transwell migration of HSC to low threshold doses of SDF-1. This phenomenon was not receptor-dependent, as agonists of membrane receptors that may bind beta2-defensin (FPRL-1), cathelicidin (CCR6) - FPRL-1 agonist, and MIP-3alpha, respectively, did not show similar priming effects. This could be explained by affected distribution of membrane lipids by cationic peptides. In support of this notion, an inhibitor of cell membrane raft formation (methyl-b-cyclodextran) inhibited the priming effect of both compounds, indicating this effect is dependent on CXCR4 incorporation into lipid rafts. Direct confocal analysis of CXCR4 and lipid raft colocalization in the presence or absence of cationic peptides confirmed these findings. Because leucopheresis products are enriched in activated granulocytes that release beta2-defensin and cathelicidin, we tested whether this may explain why mobilized peripheral blood stem cells (PBSC) engraft faster compared to HSC isolated directly from bone marrow (BM) in a murine BM transplant model. Accordingly, syngeneic BMMNCs were exposed ex vivo to beta2-defensin or cathelicidin for 30 minutes and subsequently transplanted into lethally irradiated recipients. We noted that animals transplanted with BM cells primed by those cationic peptides showed accelerated recovery of platelets and neutrophils by ∼3-5 days compared to unprimed control cells. We envision that small cationic peptides, which primarily possess antimicrobial functions and are harmless to mammalian cells, could be clinically applied to prime human HSC before transplantation. This novel approach would be particularly important in cord blood transplantation, where the number of HSC availabel for transplantation is usually limited. We postulate that this promising strategy warrants further investigations. Disclosures: No relevant conflicts of interest to declare.

scholarly journals HCMV Infection of Humanized Mice after Transplantation of G-CSF–Mobilized Peripheral Blood Stem Cells from HCMV-Seropositive Donors

2014 ◽  
Vol 20 (1) ◽  
pp. 132-135 ◽  
Author(s):  
Morgan Hakki ◽  
Devorah C. Goldman ◽  
Daniel N. Streblow ◽  
Kimberly L. Hamlin ◽  
Craig N. Krekylwich ◽  
...  
Keyword(s):  
Stem Cells ◽  
Peripheral Blood ◽  
Hcmv Infection ◽  
Humanized Mice ◽  
Peripheral Blood Stem Cells ◽  
Blood Stem Cells ◽  
Mobilized Peripheral Blood

Efficient and Durable Gene Marking of Hematopoietic Progenitor Cells in Nonhuman Primates After Nonablative Conditioning

Blood ◽  
1999 ◽  
Vol 94 (7) ◽  
pp. 2271-2286 ◽  
Author(s):  
M. Rosenzweig ◽  
T.J. MacVittie ◽  
D. Harper ◽  
D. Hempel ◽  
R.L. Glickman ◽  
...  
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Stem Cell ◽  
Peripheral Blood ◽  
Nonhuman Primates ◽  
Peripheral Blood Stem Cells ◽  
Hematopoietic Stem ◽  
Blood Stem Cells ◽  
High Level

Optimization of mobilization, harvest, and transduction of hematopoietic stem cells is critical to successful stem cell gene therapy. We evaluated the utility of a novel protocol involving Flt3-ligand (Flt3-L) and granulocyte colony-stimulating factor (G-CSF) mobilization of peripheral blood stem cells and retrovirus transduction using hematopoietic growth factors to introduce a reporter gene, murine CD24 (mCD24), into hematopoietic stem cells in nonhuman primates. Rhesus macaques were treated with Flt3-L (200 μg/kg) and G-CSF (20 μg/kg) for 7 days and autologous CD34+ peripheral blood stem cells harvested by leukapheresis. CD34+ cells were transduced with an MFGS-based retrovirus vector encoding mCD24 using 4 daily transductions with centrifugations in the presence of Flt3-L (100 ng/mL), human stem cell factor (50 ng/mL), and PIXY321 (50 ng/mL) in serum-free medium. An important and novel feature of this study is that enhanced in vivo engraftment of transduced stem cells was achieved by conditioning the animals with a low-morbidity regimen of sublethal irradiation (320 to 400 cGy) on the day of transplantation. Engraftment was monitored sequentially in the bone marrow and blood using both multiparameter flow cytometry and semi-quantitative DNA polymerase chain reaction (PCR). Our data show successful and persistent engraftment of transduced primitive progenitors capable of giving rise to marked cells of multiple hematopoietic lineages, including granulocytes, monocytes, and B and T lymphocytes. At 4 to 6 weeks posttransplantation, 47% ± 32% (n = 4) of granulocytes expressed mCD24 antigen at the cell surface. Peak in vivo levels of genetically modified peripheral blood lymphocytes approached 35% ± 22% (n = 4) as assessed both by flow cytometry and PCR 6 to 10 weeks posttransplantation. In addition, naı̈ve (CD45RA+and CD62L+) CD4+ and CD8+cells were the predominant phenotype of the marked CD3+ T cells detected at early time points. A high level of marking persisted at between 10% and 15% of peripheral blood leukocytes for 4 months and at lower levels past 6 months in some animals. A cytotoxic T-lymphocyte response against mCD24 was detected in only 1 animal. This degree of persistent long-lived, high-level gene marking of multiple hematopoietic lineages, including naı̈ve T cells, using a nonablative marrow conditioning regimen represents an important step toward the ultimate goal of high-level permanent transduced gene expression in stem cells.

Determination of peripheral blood stem cells by the Sysmex SE-9500

2001 ◽  
Vol 23 (4) ◽  
pp. 231-236 ◽  
Author(s):  
Liming Peng ◽  
Jiahui Yang ◽  
Hui Yang ◽  
Zhiyong Peng ◽  
Caigang Xu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Peripheral Blood ◽  
Peripheral Blood Stem Cells ◽  
Blood Stem Cells

scholarly journals Large-scale isolation of CD133+ progenitor cells from G-CSF mobilized peripheral blood stem cells

2003 ◽  
Vol 31 (1) ◽  
pp. 17-22 ◽  
Author(s):  
P R Gordon ◽  
T Leimig ◽  
A Babarin-Dorner ◽  
J Houston ◽  
M Holladay ◽  
...  
Keyword(s):  
Stem Cells ◽  
Progenitor Cells ◽  
Peripheral Blood ◽  
Large Scale ◽  
Peripheral Blood Stem Cells ◽  
Blood Stem Cells ◽  
Mobilized Peripheral Blood
Sign in / Sign up

Export Citation Format

Share Document