Low immunogenic endothelial cells endothelialize the Left Ventricular Assist Device

Abstract

Low haemocompatibility of left ventricular assist devices (LVAD) surfaces necessitates anticoagulative therapy. Endothelial cell (EC) seeding can support haemocompatibility, however, the availability of autologous ECs is limited. In contrast, allogeneic ECs are readily available in sufficient quantity, but HLA disparities induce harmful immune responses causing EC loss. In this study, we investigated the feasibility of using allogeneic low immunogenic ECs to endothelialize LVAD sintered inflow cannulas (SIC). To reduce the immunogenicity of ECs, we applied an inducible lentiviral vector to deliver short-hairpins RNA to silence HLA class I expression. HLA class I expression on ECs was conditionally silenced by up to 70%. Sufficient and comparable endothelialization rates were achieved with HLA-expressing or HLA-silenced ECs. Cell proliferation was not impaired by cell-to-Sintered Inflow Cannulas (SIC) contact or by silencing HLA expression. The levels of endothelial phenotypic and thrombogenic markers or cytokine secretion profiles remained unaffected. HLA-silenced ECs-coated SIC exhibited reduced thrombogenicity. In contrast to native ECs, HLA-silenced ECs showed lower cell lysis rates when exposed to allogeneic T cells or specific anti-HLA antibodies. Allogeneic HLA-silenced ECs could potentially become a valuable source for LVAD endothelialization to reduce immunogenicity and correspondingly the need for anticoagulative therapy which can entail severe side effects.

Fit-For-All iPSC-Derived Cell Therapies and Their Evaluation in Humanized Mice With NK Cell Immunity

“…Researchers have access to several molecular tools to disrupt β2M expression, and the choice depends on the outcome needed. Post-transcriptional manipulation by RNA interference (RNAi), either using small interfering RNA (siRNA) or short hairpin RNA (shRNA), stably downregulated or silenced the expression of HLA proteins in B-lymphocyte cell lines ( 58 ) in conjunction with human ESCs ( 59 ), iPSCs ( 60 ), primary human hepatocytes ( 61 ), endothelial cells ( 62 , 63 ) and in a perfusion system at the whole organ level in rat kidney ( 64 ). Silencing with RNAi proved to be stable even after the iPSCs were differentiated to megakaryocytes and even platelets ( 60 ).…”

Surface Coatings for Rotary Ventricular Assist Devices: A Systematic Review

“…Here, the included publications all showed it could significantly reduce platelet adhesion as compared with noncoated counterparts in in vitro studies. 54–56 For other coatings like amorphous carbon, 23 PEO, 27 GP IIb/IIIa receptor inhibitor, 45 silicone, 46 polyurethane, 47 Ti, 48 Ti6Al7Nb, 48 and apatite, 49 their hemocompatibility was investigated in a very limited number of publications. In the publication of UNCD, 7 the author did not investigate the hemocompatibility but gave the information by citing other publications which showed UNCD was antithrombogenic.…”

Control of Blood Coagulation by Hemocompatible Material Surfaces—A Review

“…One engineering strategy for the endothelialization of material surfaces is in vitro cell seeding, where autologous or allogenic ECs are harvested and cultured, and subsequently seeded on the blood-contacting device surface before implantation [ 217 , 218 ]. However, in vitro EC seeding is a very time-consuming and cost-intensive procedure, and is challenging because of problems with cell sourcing, cell stability, and cell viability [ 58 ].…”