title: Scalable generation of universal platelets from human induced pluripotent stem cells.
abstract: Human induced pluripotent stem cells (iPSCs) provide a potentially replenishable source for the production of transfusable platelets. Here, we describe a method to generate megakaryocytes (MKs) and functional platelets from iPSCs in a scalable manner under serum/feeder-free conditions. The method also permits the cryopreservation of MK progenitors, enabling a rapid "surge" capacity when large numbers of platelets are needed. Ultrastructural/morphological analyses show no major differences between iPSC platelets and human blood platelets. iPSC platelets form aggregates, lamellipodia, and filopodia after activation and circulate in macrophage-depleted animals and incorporate into developing mouse thrombi in a manner identical to human platelets. By knocking out the β2-microglobulin gene, we have generated platelets that are negative for the major histocompatibility antigens. The scalable generation of HLA-ABC-negative platelets from a renewable cell source represents an important step toward generating universal platelets for transfusion as well as a potential strategy for the management of platelet refractoriness.
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Feng
authors.affiliation: Department of Pharmacology, University of Illinois College of Medicine, Chicago, IL 60612, USA.
authors.forename: Namrata
authors.initials: N
authors.lastname: Shabrani
authors.affiliation: Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115.
authors.forename: Jonathan N
authors.initials: JN
authors.lastname: Thon
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Hongguang
authors.initials: H
authors.lastname: Huo
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Austin
authors.initials: A
authors.lastname: Thiel
authors.affiliation: Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115.
authors.forename: Kellie R
authors.initials: KR
authors.lastname: Machlus
authors.affiliation: Department of Pharmacology, University of Illinois College of Medicine, Chicago, IL 60612, USA.
authors.forename: Kyungho
authors.initials: K
authors.lastname: Kim
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: Brooks
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Chenmei
authors.initials: C
authors.lastname: Luo
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Erin A
authors.initials: EA
authors.lastname: Kimbrel
authors.affiliation: Allele Biotechnology, San Diego, CA 92121, USA.
authors.forename: Jiwu
authors.initials: J
authors.lastname: Wang
authors.affiliation: MacLean Hospital, Harvard Medical School, Belmont, MA 02478, USA.
authors.forename: Kwang-Soo
authors.initials: KS
authors.lastname: Kim
authors.affiliation: Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA; Harvard Medical School, Boston, MA 02115; Vascular Biology Program, Department of Surgery, Boston Children's Hospital, Boston, MA 02115, USA.
authors.forename: Joseph
authors.initials: J
authors.lastname: Italiano
authors.affiliation: Department of Pharmacology, University of Illinois College of Medicine, Chicago, IL 60612, USA.
authors.forename: Jaehyung
authors.initials: J
authors.lastname: Cho
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA.
authors.forename: Shi-Jiang
authors.initials: SJ
authors.lastname: Lu
authors.affiliation: Advanced Cell Technology, Marlborough, MA 01752, USA. Electronic address: rlanza@advancedcell.com.
authors.forename: Robert
authors.initials: R
authors.lastname: Lanza
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D055786:Gene Knockout Techniques; D006680:HLA Antigens / Q000235:genetics / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D051917:Leukosialin / Q000378:metabolism; D008297:Male; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism; D017713:Platelet Transfusion / Q000379:methods; D015203:Reproducibility of Results; D014183:Transplantation, Heterologous; D001613:beta 2-Microglobulin / Q000235:genetics / Q000378:metabolism
chemical_list: D018952:Antigens, CD34; D006680:HLA Antigens; D051917:Leukosialin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D001613:beta 2-Microglobulin