The overall goal of Project 3 is to develop a method to efficiently and reproducibly differentiate induced pluripotent stem (IPS) cells into hematopoietic stem cells (HSCs).
Aim 1 of this project will determine the best conditions for the mesodermal differentiation of iPS cells and the generation of HSCs.
This aim will test the hypothesis that an engineered human stromal cell line offers the best method to reliably differentiate IPS cells into hematopoietic precursors. Experiments will compare methods of differentiating IPS cells using embryoid bodies (EB) cultures and stromal cell lines to support the differentiation of IPS cells into HSCs. Various elements of these two differentiation methods will be studied to determine the best method to promote mesodermal differentiation and HSC creation.
The second aim i s to determine the optimal cytokine conditions for the survival, growth and expansion of HSCs generated from iPS cells.
This aim will test the hypothesis that HSCs generated from IPS are more similar in their cytokine responses to fetal HSCs than adult HSCs. Accumulation of HSCs in culture requires conditions that favor their survival and minimize their differentiation into committed progenitors. Various cytokines known to play a role in the early stages of hematopoiesis will be tested in combination to compare the similarity of HSCs generated from iPS cells to those isolated from fetal tissues, umbilical cord blood and adult peripheral blood. These experiments will optimize culture conditions for the production of HSCs.
The third aim will determine the variability in the capacity of different IPS cell lines to differentiate into HSCs.
This aim will test the hypothesis that different iPS cells lines are similarly capable to form HSCs that can generate long-term multilineage reconstitution in immunodeficient mice.
This aim will test isolated HSCs derived from multiple IPS cell lines for their capacity to provide long-term reconstitution without teratoma or leukemia formation, chromosomal abnormalities or other obvious functional deficiencies. The outcome of the experiments will be development of technology to efficiently generate transplantable HSCs from genetically corrected IPS cells.
Project 3 develops methods of generating transplantable hematopoietic stem cells from pluripotent stem cells. This supports the overall program project goals to develop new methods of treating hemoglobinopathies using genetically correcting patient cells to generate blood stem cells for autologous transplantation.
|Tan, Yu-Ting; Ye, Lin; Xie, Fei et al. (2018) Respecifying human iPSC-derived blood cells into highly engraftable hematopoietic stem and progenitor cells with a single factor. Proc Natl Acad Sci U S A 115:2180-2185|
|Togarrati, Padma Priya; Sasaki, Robson T; Abdel-Mohsen, Mohamed et al. (2017) Identification and characterization of a rich population of CD34+ mesenchymal stem/stromal cells in human parotid, sublingual and submandibular glands. Sci Rep 7:3484|
|Fomin, Marina E; Beyer, Ashley I; Muench, Marcus O (2017) Human fetal liver cultures support multiple cell lineages that can engraft immunodeficient mice. Open Biol 7:|
|Beyer, Ashley I; Muench, Marcus O (2017) Comparison of Human Hematopoietic Reconstitution in Different Strains of Immunodeficient Mice. Stem Cells Dev 26:102-112|
|Fomin, Marina E; Beyer, Ashley I; Publicover, Jean et al. (2017) Higher Serum Alanine Transaminase Levels in Male Urokinase-Type Plasminogen Activator-Transgenic Mice Are Associated With Improved Engraftment of Hepatocytes but not Liver Sinusoidal Endothelial Cells. Cell Med 9:117-125|
|Muench, Marcus O; Kapidzic, Mirhan; Gormley, Matthew et al. (2017) The human chorion contains definitive hematopoietic stem cells from the fifteenth week of gestation. Development 144:1399-1411|
|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 ?-thalassemia. Proc Natl Acad Sci U S A 113:10661-5|
|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|
|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|
|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|
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