title: Generation of induced pluripotent stem cells from CD34+ cells across blood drawn from multiple donors with non-integrating episomal vectors.
abstract: The methodology to create induced pluripotent stem cells (iPSCs) affords the opportunity to generate cells specific to the individual providing the host tissue. However, existing methods of reprogramming as well as the types of source tissue have significant limitations that preclude the ability to generate iPSCs in a scalable manner from a readily available tissue source. We present the first study whereby iPSCs are derived in parallel from multiple donors using episomal, non-integrating, oriP/EBNA1-based plasmids from freshly drawn blood. Specifically, successful reprogramming was demonstrated from a single vial of blood or less using cells expressing the early lineage marker CD34 as well as from unpurified peripheral blood mononuclear cells. From these experiments, we also show that proliferation and cell identity play a role in the number of iPSCs per input cell number. Resulting iPSCs were further characterized and deemed free of transfected DNA, integrated transgene DNA, and lack detectable gene rearrangements such as those within the immunoglobulin heavy chain and T cell receptor loci of more differentiated cell types. Furthermore, additional improvements were made to incorporate completely defined media and matrices in an effort to facilitate a scalable transition for the production of clinic-grade iPSCs.
authors: Cellular Dynamics International, Inc., Madison, Wisconsin, United States of America. amack@cellulardynamics.com
authors: Amanda A
authors: AA
authors: Mack
authors: Stacie
authors: S
authors: Kroboth
authors: Deepika
authors: D
authors: Rajesh
authors: Wen Bo
authors: WB
authors: Wang
mesh_terms: D000328:Adult; D018952:Antigens, CD34 / Q000378:metabolism*; D001782:Blood Donors; D001800:Blood Specimen Collection; D049109:Cell Proliferation; D002469:Cell Separation; D065150:Cellular Reprogramming; D005260:Female; D005822:Genetic Vectors / Q000235:genetics*; D019650:Hematopoietic Stem Cell Mobilization; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007202:Indicators and Reagents; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D010957:Plasmids / Q000235:genetics*; D014162:Transfection; D019076:Transgenes / Q000235:genetics
chemical_list: D018952:Antigens, CD34; D007202:Indicators and Reagents