Human Parthenogenetic Embryonic Stem Cell-Derived Neural Stem Cells Express HLA-G and Show Unique Resistance to NK Cell-Mediated Killing

Abstract

Parent-of-origin imprints have been implicated in the regulation of neural differentiation and brain development. Previously we have shown that, despite the lack of a paternal genome, human parthenogenetic (PG) embryonic stem cells (hESCs) can form proliferating neural stem cells (NSCs) that are capable of differentiation into physiologically functional neurons while maintaining allele-specific expression of imprinted genes. Since biparental ("normal") hESC-derived NSCs (N NSCs) are targeted by immune cells, we characterized the immunogenicity of PG NSCs. Flow cytometry and immunocytochemistry revealed that both N NSCs and PG NSCs exhibited surface expression of human leukocyte antigen (HLA) class I but not HLA-DR molecules. Functional analyses using an in vitro mixed lymphocyte reaction assay resulted in less proliferation of peripheral blood mononuclear cells (PBMC) with PG compared with N NSCs. In addition, natural killer (NK) cells cytolyzed PG less than N NSCs. At a molecular level, expression analyses of immune regulatory factors revealed higher HLA-G levels in PG compared with N NSCs. In line with this finding, MIR152, which represses HLA-G expression, is less transcribed in PG compared with N cells. Blockage of HLA-G receptors ILT2 and KIR2DL4 on natural killer cell leukemia (NKL) cells increased cytolysis of PG NSCs. Together this indicates that PG NSCs have unique immunological properties due to elevated HLA-G expression.

Mouse Parthenogenetic Embryonic Stem Cells with Biparental-Like Expression of Imprinted Genes Generate Cortical-Like Neurons That Integrate into the Injured Adult Cerebral Cortex

“…For example, a collection of human Pg‐ESC lines established from 55 oocyte donors would perfectly match the three HLA loci for 80% of the Japanese population . Furthermore, the neural derivatives of Pg‐ESCs are more resistant to natural killer cells than Bp‐ESC derivatives . Mouse and human Pg‐ESCs can be differentiated in vitro and in vivo into derivatives of all germ lineages , including neural lineages .…”

“…Furthermore, the neural derivatives of Pg‐ESCs are more resistant to natural killer cells than Bp‐ESC derivatives . Mouse and human Pg‐ESCs can be differentiated in vitro and in vivo into derivatives of all germ lineages , including neural lineages . Pg‐ESCs have therefore emerged as a potent source of histocompatible cells for liver , heart , hematopoietic tissue , tendon , and the substantia nigra .…”

Using an in vitro xenoantibody-mediated complement-dependent cytotoxicity model to evaluate the complement inhibitory activity of the peptidic C3 inhibitor Cp40

“…The percentage of PI-positive cells (compared to total cells) was used to evaluate the degree of cell death. The inhibition percentage was calculated as described elsewhere [7,21,22], using the following equation: % inhibition = {1 – (A – C) / (B – C)} × 100%, where A, B, and C represent the percentage of cell death when incubated with 20% serum pretreated with Cp40 (A), 20% serum without any treatment (B), and 20% heat-inactivated serum (C), respectively.…”

Non‐coding RNAs: emerging regulatory factors in the derivation and differentiation of mammalian parthenogenetic embryonic stem cells

“…To date, PESCs have been successfully derived from several mammalian species including human, mouse, bovine, porcine, and rabbit (Bevacqua et al, 2011;De Sousa et al, 2016;Intawicha et al, 2016;Kim et al, 2016;Zhong et al, 2016). (Onodera et al, 2010;Espejel et al, 2014;Schmitt et al, 2015;Sagi et al, 2016). They have been widely used in regenerative medicine due to their capacity to differentiate into various cell types both in vitro and in vivo, and are also the most ideal material in being to study the distinct roles of paternal and maternal genomes during development of multiple systems/organs/tissues, including and neural system, brain, lung, liver, and heart, etc.…”

“…They have been widely used in regenerative medicine due to their capacity to differentiate into various cell types both in vitro and in vivo, and are also the most ideal material in being to study the distinct roles of paternal and maternal genomes during development of multiple systems/organs/tissues, including and neural system, brain, lung, liver, and heart, etc. (Onodera et al, 2010;Espejel et al, 2014;Schmitt et al, 2015;Sagi et al, 2016). Mammalian PESCs are an important way to produce patient-specific stem cell lines or terminal differentiated cells, which are histocompatible cells, for cellbased therapy.…”