Immunological properties of parthenogenetic stem cell derived cardiomyocytes and their application in cardiac tissue engineering

by Satish Galla
Doctoral thesis
Date of Examination:2016-06-14
Date of issue:2016-06-20
Advisor:Prof. Dr. Wolfram-Hubertus Zimmermann
Referee:Prof. Dr. Ralf Dressel
Referee:Prof. Dr. Michael P. Schön
Referee:Prof. Dr. Susanne Lutz
Referee:Prof. Dr. Hubertus Jarry
Referee:Prof. Dr. Thomas Meyer
Persistent Address: http://dx.doi.org/10.53846/goediss-5699

 

 

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Abstract

English

Cardiovascular diseases and their long term consequences constitute serious health and economic burdens. Although promising new therapeutic approaches to ameliorate the detrimental consequences have been introduced, there is still no therapy that would lead to a therapeutically efficient replacement of lost cardiomyocytes (CMs), e.g. after a myocardial infarction. CMs can be reliably generated from pluripotent stem cells, including embryonic and induced pluripotent stem cells. Parthenogenetic stem cells (PSCs) have recently been introduced as an alternative, attractive pluripotent stem cell entity. PSCs generated from pharmacologically activated unfertilized oocytes contain only maternal chromosomes, and show a growth and differentiation behavior similar to embryonic stem cells (ESCs). The unique chromosomal constitution of PSCs makes them largely haploidentical, creating the possibility for immunological matching. In this study we demonstrated that transgenic major histocompatibility complex (MHC)-haploidentical PSCs (H-2d/d) can be generated carrying a neomycin resistance gene (NeoR) transcribed under the control of the CM-restricted alpha myosin heavy chain (MYH6) promoter for antibiotic selection of PSC-CMs. Differentiation and cardiomyocytes purification could be achieved at a large scale in suspension culture spinner flasks. PSC-derived non-myocytes (NM) express MHC-I which was significantly upregulated after interferon gamma (IFNγ) stimulation. PSC-derived CM did not express MHC-I and MHC-II under basal conditions, but were up-regulated after IFNγ treatment. Expression of co-stimulatory molecules (CD40, CD80 and CD86) was not evident in PSC-derivatives under baseline or IFNγ stimulated conditions. PD-L1 expression was upregulated after IFNγ stimulation. In vitro immune cell proliferation assays showed that PSC-CM and PSC-NM had strong effect on lymphocyte activation, while PSC-EHM had negligible effect on lymphocyte stimulation. Implantation of PSC-CM under the kidney capsule suggested enhanced survival under MHC-matching. In conclusion, this study provides insight into the immunological properties of PSC-derived cardiomyocytes and their derivatives.
Keywords: parthenogenetic stem cells; haploidentity; cardiomyocytes; major histocompatibility complex
Schlagwörter: stem cells
 
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