PMID- 26218149 OWN - NLM STAT- MEDLINE DCOM- 20160916 LR - 20220321 IS - 1937-335X (Electronic) IS - 1937-3341 (Print) IS - 1937-3341 (Linking) VI - 21 IP - 19-20 DP - 2015 Oct TI - HLA Class I Depleted hESC as a Source of Hypoimmunogenic Cells for Tissue Engineering Applications. PG - 2559-71 LID - 10.1089/ten.TEA.2015.0105 [doi] AB - BACKGROUND: Rapidly improving protocols for the derivation of autologous cells from stem cell sources is a welcome development. However, there are many circumstances when off-the-shelf universally immunocompatible cells may be needed. Embryonic stem cells (ESCs) provide a unique opportunity to modify the original source of differentiated cells to minimize their rejection by nonautologous hosts. HYPOTHESIS: Immune rejection of nonautologous human embryonic stem cell (hESC) derivatives can be reduced by downregulating human leukocyte antigen (HLA) class I molecules, without affecting the ability of these cells to differentiate into specific lineages. METHODS AND RESULTS: Beta-2-microglobulin (B2M) expression was decreased by lentiviral transduction using human anti-HLA class I light-chain B2M short hairpin RNA. mRNA levels of B2M were decreased by 90% in a RUES2-modified hESC line, as determined by quantitative real time-polymerase chain reaction analysis. The transduced cells were selected under puromycin pressure and maintained in an undifferentiated state. The latter was confirmed by Oct4 and Nanog expression, and by the formation of characteristic round-shaped colonies. B2M downregulation led to diminished HLA-I expression on the cell surface, as determined by flow cytometry. When used as target cells in a mixed lymphocyte reaction assay, transduced hESCs and their differentiated derivatives did not stimulate allogeneic T-cell proliferation. Using a cardiac differentiation protocol, transduced hESCs formed a confluent layer of cardiac myocytes and maintained a low level of B2M expression. Transduced hESCs were also successfully differentiated into a hepatic lineage, validating their capacity to differentiate into multiple lineages. CONCLUSIONS: HLA-I depletion does not preclude hESC differentiation into cardiac or hepatic lineages. This methodology can be used to engineer tissue from nonautologous hESC sources with improved immunocompatibility. FAU - Karabekian, Zaruhi AU - Karabekian Z AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. AD - 2 L.A.Orbeli Institute of Physiology, National Academy of Sciences , Yerevan, Armenia . FAU - Ding, Hao AU - Ding H AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. FAU - Stybayeva, Gulnaz AU - Stybayeva G AD - 3 Department of Biomedical Engineering, University of California Davis , Davis, California. FAU - Ivanova, Irina AU - Ivanova I AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. FAU - Muselimyan, Narine AU - Muselimyan N AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. FAU - Haque, Amranul AU - Haque A AD - 3 Department of Biomedical Engineering, University of California Davis , Davis, California. FAU - Toma, Ian AU - Toma I AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. FAU - Posnack, Nikki G AU - Posnack NG AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. FAU - Revzin, Alexander AU - Revzin A AD - 3 Department of Biomedical Engineering, University of California Davis , Davis, California. FAU - Leitenberg, David AU - Leitenberg D AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. FAU - Laflamme, Michael A AU - Laflamme MA AD - 4 Institute for Stem Cell and Regenerative Medicine, Center for Cardiovascular Biology, University of Washington , Seattle, Washington. FAU - Sarvazyan, Narine AU - Sarvazyan N AD - 1 Pharmacology and Physiology Department, School of Medicine and Health Sciences, The George Washington University , Washington, District of Columbia. LA - eng GR - R21 HL122882/HL/NHLBI NIH HHS/United States PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150910 PL - United States TA - Tissue Eng Part A JT - Tissue engineering. Part A JID - 101466659 RN - 0 (Histocompatibility Antigens Class I) SB - IM MH - Cell Differentiation/physiology MH - Cell Line MH - Cell Proliferation/physiology MH - Embryonic Stem Cells/*cytology/metabolism MH - Flow Cytometry MH - Histocompatibility Antigens Class I/metabolism MH - Humans MH - Immunohistochemistry MH - Pluripotent Stem Cells/*cytology/metabolism MH - Tissue Engineering/*methods PMC - PMC4605353 EDAT- 2015/07/29 06:00 MHDA- 2016/09/17 06:00 PMCR- 2016/10/01 CRDT- 2015/07/29 06:00 PHST- 2015/07/29 06:00 [entrez] PHST- 2015/07/29 06:00 [pubmed] PHST- 2016/09/17 06:00 [medline] PHST- 2016/10/01 00:00 [pmc-release] AID - 10.1089/ten.tea.2015.0105 [pii] AID - 10.1089/ten.TEA.2015.0105 [doi] PST - ppublish SO - Tissue Eng Part A. 2015 Oct;21(19-20):2559-71. doi: 10.1089/ten.TEA.2015.0105. Epub 2015 Sep 10.