The purpose of Core B is to provide critical support for molecular/cellular analyses, ceil and vector banking, and generation of animal-product-free reagents fbr research needs performed under the three Projects and Core C. The specific service aims of Core B are:
Aim 1 : To provide cellular analysis of iPS and other cell lines. Core B will be responsible for providing expertise, assistance and coordination of immunohistochemical staining for expression antigens in iPS and differentiated cell lines, and for analysis of karyotype.
Aim 2 : To provide molecular analysis of iPS and other cell lines. Core B will provide molecular analyses of cell lines to validate their status as IPS, primary, or differentiated cell lines. This will include gene expression profiles, promoter methylation, and microRNA analyses. The core will be responsible for confirming B-globin genotype for mutant and corrected cell lines and microsatellite fingerprint profiles. One goal is to define a molecular expression profile or signature to monitor and validate in vitro differentiation of IPS to hematopoietic precursor cells competent to repopulate bone marrow in vivo.
Aim 3 : To provide cell and vector banking and evaluate cells for eventual therapeutic development. In anticipation that materials produced may be transitioned to the clinic, we will establish a centralized location and database to document and maintain key ceil lines, vectors, reagents, and other ancillary materials. This will be done in conjunction with Core A.
Aim 4 : To engineer a human cell line to replace mouse 0P9 stromal cells used for differentiation of IPS cells into hematopoietic cell lines. One of the most effective methods used to date has employed co-culture of IPS and hES cells with the mouse stromal 0P9 cell line. Since exposure to animal derived products might introduce pathogens with potential adverse safety consequences for patients, development of a human stromal cell line to replace 0P9 will be one component of creating animal-product free processes for generating gene-corrected HSCs.
Aim 5 : To perform routine reprogramming of primary cell lines into IPS ceil lines for use in early stage research for projects 2 and 3. Somatic cells will be reprogrammed to IPS cells for the gene connection (Project 2) and HCS differentiation (Project 3) studies will initially be performed using retroviral vectors carrying 4 reprogramming genes (0CT4, S0X2, KLF4, and cMYC). As new reprogramming technologies are developed in Project 1 they will be applied for the generation of IPS cells.

Public Health Relevance

This core provides services critical to accomplishing the research goals of the overall program project that aims to develop new methods of treating hemoglobinopathies sickle cell anemia and beta-thalassemia using genetically corrected patient cells to generate hematopoietic stem cells for autologous transplantation, improving these therapies would significantly improve the quality of life among afflicted individuals and decrease the social and economic Burden that they impose on individuals and the healthcare system.

National Institute of Health (NIH)
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Research Program Projects (P01)
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University of California San Francisco
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Suzuki, Shingo; Sargent, R Geoffrey; Illek, Beate et al. (2016) TALENs Facilitate Single-step Seamless SDF Correction of F508del CFTR in Airway Epithelial Submucosal Gland Cell-derived CF-iPSCs. Mol Ther Nucleic Acids 5:e273
Beyer, Ashley I; Muench, Marcus O (2016) Comparison of human hematopoietic reconstitution in different strains of immunodeficient mice. Stem Cells Dev :
Ye, Lin; Wang, Jiaming; Tan, Yuting et al. (2016) Genome editing using CRISPR-Cas9 to create the HPFH genotype in HSPCs: An approach for treating sickle cell disease and β-thalassemia. Proc Natl Acad Sci U S A 113:10661-5
Baimukanova, Gyulnar; Miyazawa, Byron; Potter, Daniel R et al. (2016) Platelets regulate vascular endothelial stability: assessing the storage lesion and donor variability of apheresis platelets. Transfusion 56 Suppl 1:S65-75
Wiemels, J L; de Smith, A J; Xiao, J et al. (2016) A functional polymorphism in the CEBPE gene promoter influences acute lymphoblastic leukemia risk through interaction with the hematopoietic transcription factor Ikaros. Leukemia 30:1194-7
Mahajan, Maya M; Cheng, Betty; Beyer, Ashley I et al. (2015) A quantitative assessment of the content of hematopoietic stem cells in mouse and human endosteal-bone marrow: a simple and rapid method for the isolation of mouse central bone marrow. BMC Hematol 15:9
Muench, Marcus O; Beyer, Ashley I; Fomin, Marina E et al. (2014) The adult livers of immunodeficient mice support human hematopoiesis: evidence for a hepatic mast cell population that develops early in human ontogeny. PLoS One 9:e97312
Xie, Fei; Ye, Lin; Chang, Judy C et al. (2014) Seamless gene correction of β-thalassemia mutations in patient-specific iPSCs using CRISPR/Cas9 and piggyBac. Genome Res 24:1526-33
Ye, Lin; Wang, Jiaming; Beyer, Ashley I et al. (2014) Seamless modification of wild-type induced pluripotent stem cells to the natural CCR5Δ32 mutation confers resistance to HIV infection. Proc Natl Acad Sci U S A 111:9591-6
Fomin, M E; Togarrati, P P; Muench, M O (2014) Progress and challenges in the development of a cell-based therapy for hemophilia A. J Thromb Haemost 12:1954-65

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