scholarly journals Evaluation of the Spanish population coverage of a prospective HLA haplobank of induced pluripotent stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Belén Álvarez-Palomo ◽  
Iris García-Martinez ◽  
Jorge Gayoso ◽  
Angel Raya ◽  
Anna Veiga ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Spanish Population ◽  
Population Coverage ◽  
Advanced Therapy ◽  
Blood Banks ◽  
Induced Pluripotent ◽  
Cord Blood Banks

Abstract Background iPSC (induced pluripotent stem cells) banks of iPSC lines with homozygous HLA (human leukocyte antigen) haplotypes (haplobanks) are proposed as an affordable and off-the-shelf approach to allogeneic transplantation of iPSC derived cell therapies. Cord blood banks offer an extensive source of HLA-typed cells suitable for reprogramming to iPSC. Several initiatives worldwide have been undertaken to create national and international iPSC haplobanks that match a significant part of a population. Methods To create an iPSC haplobank that serves the Spanish population (IPS-PANIA), we have searched the Spanish Bone Marrow Donor Registry (REDMO) to identify the most frequently estimated haplotypes. From the top ten donors identified, we estimated the population coverage using the criteria of zero mismatches in HLA-A, HLA-B, and HLA-DRB1 with different stringencies: high resolution, low resolution, and beneficial mismatch. Results We have calculated that ten cord blood units from homozygous donors stored at the Spanish cord blood banks can provide HLA-A, HLA-B, and HLA-DRB1 matching for 28.23% of the population. Conclusion We confirm the feasibility of using banked cord blood units to create an iPSC haplobank that will cover a significant percentage of the Spanish and international population for future advanced therapy replacement strategies.

scholarly journals Evaluation of the Spanish Population Coverage of a Prospective HLA Haplobank of Induced Pluripotent Stem Cells.

2020 ◽  
Author(s):  
Belen Alvarez-Palomo ◽  
Iris Garcia-Martinez ◽  
Jorge Gayoso ◽  
Angel Raya ◽  
Anna Veiga ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Spanish Population ◽  
Population Coverage ◽  
Advanced Therapy ◽  
Blood Banks ◽  
Induced Pluripotent ◽  
Cord Blood Banks

Abstract Background iPSC (Induced Pluripotent Stem Cells) banks of iPSC lines with homozygous HLA (Human Leukocyte Antigen) haplotypes (haplobanks) are proposed as an affordable and off-the-shelf approach to allogeneic transplantation of iPSC derived cell therapies. Cord blood banks offer an extensive source of HLA-typed cells suitable for reprogramming to iPSC. Several initiatives worldwide have been undertaken to create national and international iPSC haplobanks that match a significant part of a population. Methods To create an iPSC haplobank that serves the Spanish population (IPS-PANIA), we have searched the Spanish Bone Marrow Donor Registry (REDMO) to identify the most frequently estimated haplotypes. From the top ten donors identified, we estimated the population coverage using the criteria of zero mismatches in HLA-A, -B and -DRB1 with different stringencies: high resolution, low resolution and beneficial mismatch. Results We have calculated that ten cord blood units from homozygous donors stored at the Spanish cord blood banks can provide a HLA-A, HLA-B and HLA-DRB1 matching for 28,23% of the population. Conclusion We confirm the feasibility of using banked cord blood units to create an iPSC haplobank that will cover a significant percentage of the Spanish and international population for future advanced therapy replacement strategies.

Genetic reprogramming of cord blood derived endothelial colony forming cells towards human induced pluripotent stem cells using episomal plasmids

2021 ◽  
pp. 1-18
Author(s):  
Osama Shahid ◽  
Sumbul Shamim ◽  
Jahan ara Ainuddin ◽  
Mohsin Wahid
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
University Hospital ◽  
Human Umbilical Cord Blood ◽  
Human Umbilical Cord ◽  
Time Duration ◽  
Endothelial Colony Forming Cells ◽  
Induced Pluripotent

Abstract Objective: This study aimed to isolate human umbilical cord blood derived endothelial colony forming cells (ECFCs) followed by their integration free reprogramming towards induced pluripotent stem cells (iPSCs) and molecular characterization of both cell types using multicolor flowcytometery and immunofluorescence respectively. Methods: The cord blood was collected from 37-39 weeks of gestational ages after C-section ex-utero from Dow University Hospital. The ECFCs isolated after ficoll based separation of cord blood mononuclear cells (CBMNCs) which on emergence characterized through flow cytometry and reprogrammed towards induced pluripotent stem cells (iPSCs) using episomal vectors, the iPSCs were characterized using immunofluorescence. The study was conducted at Stem Cells and Regenerative lab, Dow Research Institute of Biotechnology and Biomedical Sciences, Dow University of health sciences OJHA campus. The study time duration was about one year (October 2017-October 2018), study design was in vitro experimental. The sample size of the study was n=3.   Results: The isolated ECFCs were evaluated using Flowcytometery which showed positive expression for CD31, CD34, CD146 cell surface markers and negative for CD90. The successful reprogramming of ECFCs towards iPSCs was confirmed by immunofluorescence using OCT-4 which is considered to be a master regulator of pluripotency.  Conclusion: To the best of our knowledge this study was the first attempt to integration free reprogramming of cord blood derived endothelial colony forming cells towards induced pluripotent stem using Episomal plasmids. Cells that have been isolated from cord blood and those that have been reprogrammed both have potential therapeutic applications in regenerative medicine. Continuous...

Patient-Derived Induced Pluripotent Stem Cells Recapitulate Hematopoietic Abnormalities of Juvenile Myelomonocytic Leukemia

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 637-637
Author(s):  
Gandre-Babbe Shilpa ◽  
Stella T Chou ◽  
Deborah L. French ◽  
Michelle Kang ◽  
Julie Weng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Vector System ◽  
Juvenile Myelomonocytic Leukemia ◽  
Malignant Cells ◽  
Gm Csf ◽  
Induced Pluripotent ◽  
Myelomonocytic Leukemia

Abstract Abstract 637 Juvenile Myelomonocytic Leukemia (JMML) is an aggressive childhood myeloproliferative disorder caused by NF1, NRAS, KRAS, PTPN11 or CBL gene mutations that induce Ras pathway activation with associated hypersensitivity to cytokine stimulation in myeloid progenitor cells. Understanding the pathophysiology of JMML and developing new treatments is constrained by limited access to relevant patient material. To address this problem, we generated induced pluripotent stem cells (iPSCs) from normal neonatal umbilical cord blood and two JMML patients with different mutations: 1) a somatic heterozygous E76K substitution in PTPN11 and 2) a CBL Y371H substitution arising from a germline mutation. We created iPSCs from the patient's heterozygous CBL Y371H newborn cord blood cells prior to the diagnosis of JMML and then from peripheral blood at age 7 months, when JMML ensued with outgrowth of CBL Y371H homozygous malignant cells. We reprogrammed control and malignant cells using the STEMCCA lentiviral vector system that expresses OCT4, KLF4, MYC and SOX2 in a doxycyline-inducible fashion. Resultant iPSCs exhibited hallmark features after more than twenty passages, including characteristic morphology, expression of endogenous pluripotency markers, silencing of viral reprogramming genes in the absence of doxycycline, normal karyotype and formation of endoderm-, ectoderm- and mesoderm-derived tissues in teratoma assays. Relevant PTPN11 and CBL genotypes of these clones were confirmed by DNA sequencing. Selected iPSC clones were differentiated into blood by inducing the formation of embryoid bodies in serum free medium with defined cytokines. By day 8–9, hematopoietic progenitors (CD43+, CD235+, CD41+) with erythroid, megakaryocytic and myeloid potential developed from both control (n = 2 lines) and JMML iPSCs (n= 2 lines from each patient). In methylcellulose colony assays, JMML progenitors exhibited relative hypersensitivity to GM-CSF, as reflected by increased numbers and larger size of myeloid colonies at limiting GM-CSF concentrations. In liquid cultures containing SCF, TPO, EPO, IL-3, IL-11 and IGF-1, with or without GM-CSF, the JMML progenitors produced increased proportions of CD33+CD14+ myelomonocytic cells compared to controls. Moreover, in GM-CSF dose-response assays, single cell phospho-flow cytometry analysis showed sustained STAT5 activation in JMML myeloid cells vs. controls. Thus, key pathological features of JMML, including propensity to myelomonocytic cell expansion and GM-CSF hypersensitivity, are recapitulated by in vitro differentiation of JMML iPSCs. Our findings illustrate the utility of iPSCs for modeling human blood disorders and more specifically, provide renewable sources of biologically relevant, patient-derived cells in which to explore the pathophysiology and therapy of JMML. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Functional differentiation of midbrain neurons from human cord blood-derived induced pluripotent stem cells

2014 ◽  
Vol 5 (2) ◽  
pp. 35 ◽  
Author(s):  
Nancy Stanslowsky ◽  
Alexandra Haase ◽  
Ulrich Martin ◽  
Maximilian Naujock ◽  
Andreas Leffler ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Functional Differentiation ◽  
Human Cord Blood ◽  
Midbrain Neurons ◽  
Induced Pluripotent
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