PMID- 32439978
OWN - NLM
STAT- MEDLINE
DCOM- 20201201
LR  - 20210521
IS  - 2045-2322 (Electronic)
IS  - 2045-2322 (Linking)
VI  - 10
IP  - 1
DP  - 2020 May 21
TI  - Generation and characterization of HLA-universal platelets derived from induced 
      pluripotent stem cells.
PG  - 8472
LID - 10.1038/s41598-020-65577-x [doi]
LID - 8472
AB  - Platelet demand has increased around the world. However, the inadequacy of 
      donors, the risk of transfusion-transmitted infections and associated reactions, 
      and the refractory nature of platelet transfusions are among the limitations of 
      allogeneic platelet transfusions. To alleviate these problems, we propose 
      generating platelets in a laboratory that do not induce alloimmunity to human 
      leukocyte antigen (HLA) class I, which is a major cause of immune reaction in 
      platelet transfusion refractoriness. Induced pluripotent stem cells (iPSCs) were 
      generated from peripheral blood mononuclear cells (PBMCs) of a healthy Thai 
      woman. We then knocked out the beta2-microglobulin (beta2m) gene in the cells using 
      paired CRISPR/Cas9 nickases and sequentially differentiated the cells into 
      haematopoietic stem cells (HSCs), megakaryocytes (MKs) and platelets. Silencing 
      of HLA class I expression was observed on the cell surface of beta2m-knockout iPSCs, 
      iPSC-derived HSCs, MKs and platelets. The HLA-universal iPSC-derived platelets 
      were shown to be activated, and they aggregated after stimulation. In addition, 
      our in vivo platelet survival experiments demonstrated that human platelets were 
      detectable at 2 and 24 hours after injecting the beta2m-KO MKs. In summary, we 
      successfully generated functional iPSC-derived platelets in vitro without HLA 
      class I expression by knocking out the beta2m gene using paired CRISPR/Cas9 
      nickases.
FAU - Norbnop, Phatchara
AU  - Norbnop P
AD  - Doctor of Philosophy Program in Medical Sciences, Faculty of Medicine, 
      Chulalongkorn University, Bangkok, 10330, Thailand.
AD  - Center of Excellence for Medical Genomics, Department of Pediatrics, Faculty of 
      Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
FAU - Ingrungruanglert, Praewphan
AU  - Ingrungruanglert P
AD  - Stem Cell and Cell Therapy Research Unit, Faculty of Medicine, Chulalongkorn 
      University, Bangkok, 10330, Thailand.
AD  - Department of Pharmacology, Faculty of Medicine, Chulalongkorn University, 
      Bangkok, 10330, Thailand.
FAU - Israsena, Nipan
AU  - Israsena N
AD  - Stem Cell and Cell Therapy Research Unit, Faculty of Medicine, Chulalongkorn 
      University, Bangkok, 10330, Thailand.
AD  - Department of Pharmacology, Faculty of Medicine, Chulalongkorn University, 
      Bangkok, 10330, Thailand.
FAU - Suphapeetiporn, Kanya
AU  - Suphapeetiporn K
AD  - Center of Excellence for Medical Genomics, Department of Pediatrics, Faculty of 
      Medicine, Chulalongkorn University, Bangkok, 10330, Thailand. 
      kanya.su@chula.ac.th.
AD  - Excellence Center for Genomics and Precision Medicine, King Chulalongkorn 
      Memorial Hospital, the Thai Red Cross Society, Bangkok, 10330, Thailand. 
      kanya.su@chula.ac.th.
FAU - Shotelersuk, Vorasuk
AU  - Shotelersuk V
AD  - Center of Excellence for Medical Genomics, Department of Pediatrics, Faculty of 
      Medicine, Chulalongkorn University, Bangkok, 10330, Thailand.
AD  - Excellence Center for Genomics and Precision Medicine, King Chulalongkorn 
      Memorial Hospital, the Thai Red Cross Society, Bangkok, 10330, Thailand.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20200521
PL  - England
TA  - Sci Rep
JT  - Scientific reports
JID - 101563288
RN  - 0 (Histocompatibility Antigens Class I)
SB  - IM
MH  - Animals
MH  - Blood Platelets/*cytology/metabolism
MH  - Cell Differentiation
MH  - Female
MH  - Hematopoietic Stem Cells/*cytology/metabolism
MH  - Histocompatibility Antigens Class I/*metabolism
MH  - Humans
MH  - Induced Pluripotent Stem Cells/*cytology/metabolism
MH  - Leukocytes, Mononuclear/*cytology/metabolism
MH  - Megakaryocytes/*cytology/metabolism
MH  - Mice
MH  - Mice, Inbred NOD
MH  - Mice, SCID
PMC - PMC7242456
COIS- The authors declare no competing interests.
EDAT- 2020/05/23 06:00
MHDA- 2020/12/02 06:00
PMCR- 2020/05/21
CRDT- 2020/05/23 06:00
PHST- 2020/02/14 00:00 [received]
PHST- 2020/04/22 00:00 [accepted]
PHST- 2020/05/23 06:00 [entrez]
PHST- 2020/05/23 06:00 [pubmed]
PHST- 2020/12/02 06:00 [medline]
PHST- 2020/05/21 00:00 [pmc-release]
AID - 10.1038/s41598-020-65577-x [pii]
AID - 65577 [pii]
AID - 10.1038/s41598-020-65577-x [doi]
PST - epublish
SO  - Sci Rep. 2020 May 21;10(1):8472. doi: 10.1038/s41598-020-65577-x.