HLA Class I depleted hESC as a source of hypoimmunogenic cells for tissue engineering applications

Zaruhi Karabekian, Hao Ding, Gulnaz Stybayeva, Irina Ivanova, Narine Muselimyan, Amranul Haque, Ian Toma, Nikki G. Posnack, Alexander Revzin, David Leitenberg, Michael A. Laflamme, Narine Sarvazyan

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)2559-2571
Number of pages13
JournalTissue Engineering - Part A
Volume21
Issue number19-20
DOIs
StatePublished - Oct 1 2015

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Cell Engineering
beta 2-Microglobulin
Tissue Engineering
Antigens
HLA Antigens
Stem cells
Tissue engineering
Down-Regulation
Derivatives
Puromycin
T-cells
Lymphocytes
Flow cytometry
Polymerase chain reaction
Cell proliferation
Mixed Lymphocyte Culture Test
Histocompatibility
RNA
Liver
Small Interfering RNA

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biomedical Engineering
  • Biomaterials

Cite this

Karabekian, Z., Ding, H., Stybayeva, G., Ivanova, I., Muselimyan, N., Haque, A., ... Sarvazyan, N. (2015). HLA Class I depleted hESC as a source of hypoimmunogenic cells for tissue engineering applications. Tissue Engineering - Part A, 21(19-20), 2559-2571. https://doi.org/10.1089/ten.tea.2015.0105

HLA Class I depleted hESC as a source of hypoimmunogenic cells for tissue engineering applications. / Karabekian, Zaruhi; Ding, Hao; Stybayeva, Gulnaz; Ivanova, Irina; Muselimyan, Narine; Haque, Amranul; Toma, Ian; Posnack, Nikki G.; Revzin, Alexander; Leitenberg, David; Laflamme, Michael A.; Sarvazyan, Narine.

In: Tissue Engineering - Part A, Vol. 21, No. 19-20, 01.10.2015, p. 2559-2571.

Research output: Contribution to journalArticle

Karabekian, Z, Ding, H, Stybayeva, G, Ivanova, I, Muselimyan, N, Haque, A, Toma, I, Posnack, NG, Revzin, A, Leitenberg, D, Laflamme, MA & Sarvazyan, N 2015, 'HLA Class I depleted hESC as a source of hypoimmunogenic cells for tissue engineering applications', Tissue Engineering - Part A, vol. 21, no. 19-20, pp. 2559-2571. https://doi.org/10.1089/ten.tea.2015.0105
Karabekian Z, Ding H, Stybayeva G, Ivanova I, Muselimyan N, Haque A et al. HLA Class I depleted hESC as a source of hypoimmunogenic cells for tissue engineering applications. Tissue Engineering - Part A. 2015 Oct 1;21(19-20):2559-2571. https://doi.org/10.1089/ten.tea.2015.0105
Karabekian, Zaruhi ; Ding, Hao ; Stybayeva, Gulnaz ; Ivanova, Irina ; Muselimyan, Narine ; Haque, Amranul ; Toma, Ian ; Posnack, Nikki G. ; Revzin, Alexander ; Leitenberg, David ; Laflamme, Michael A. ; Sarvazyan, Narine. / HLA Class I depleted hESC as a source of hypoimmunogenic cells for tissue engineering applications. In: Tissue Engineering - Part A. 2015 ; Vol. 21, No. 19-20. pp. 2559-2571.
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abstract = "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.",
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AU - Ding, Hao

AU - Stybayeva, Gulnaz

AU - Ivanova, Irina

AU - Muselimyan, Narine

AU - Haque, Amranul

AU - Toma, Ian

AU - Posnack, Nikki G.

AU - Revzin, Alexander

AU - Leitenberg, David

AU - Laflamme, Michael A.

AU - Sarvazyan, Narine

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