Transplantation of MHC-mismatched mouse embryonic stem cell-derived thymic epithelial progenitors and MHC-matched bone marrow prevents autoimmune diabetes

Abstract

Background Type 1 diabetes (T1D) is an autoimmune disease resulting from the destruction of insulin-secreting islet β cells by autoreactive T cells. Non-obese diabetic (NOD) mice are the widely used animal model for human T1D. Autoimmunity in NOD mice is associated with particular major histocompatibility complex (MHC) loci and impaired islet autoantigen expression and/or presentation in the thymus, which results in defects in both central and peripheral tolerance. It has been reported that induction of mixed chimerism with MHC-mismatched, but not MHC-matched donor bone marrow (BM) transplants prevents the development T1D in NOD mice. We have reported that mouse embryonic stem cells (mESCs) can be selectively induced in vitro to generate thymic epithelial progenitors (TEPs) that further develop into thymic epithelial cells (TECs) in vivo to support T cell development. Methods To determine whether transplantation of MHC-mismatched mESC-TEPs could prevent the development of insulitis and T1D, NOD mice were conditioned and injected with MHC-mismatched B6 mESC-TEPs and MHC-matched BM from H-2 g7 B6 mice. The mice were monitored for T1D development. The pancreas, spleen, BM, and thymus were then harvested from the mice for evaluation of T1D, insulitis, chimerism levels, and T cells. Results Transplantation of MHC-mismatched mESC-TEPs and MHC-matched donor BM prevented insulitis and T1D development in NOD mice. This was associated with higher expression of proinsulin 2, a key islet autoantigen in the mESC-TECs, and an increased number of regulatory T cells. Conclusions Our results suggest that embryonic stem cell-derived TEPs may offer a new approach to control T1D. Electronic supplementary material The online version of this article (10.1186/s13287-019-1347-1) contains supplementary material, which is available to authorized users.

Embryonic Trophectoderm Secretomics Reveals Chemotactic Migration and Intercellular Communication of Endometrial and Circulating MSCs in Embryonic Implantation

“…We aimed to build an in vitro model of cell-to-cell communication through EVs and soluble proteins. To demonstrate the functional transfer between embryonic trophectoderm and maternal MSC of protein-containing secretome, we first confirmed that EVs isolated from conditioned media of BBT cells previously labeled with PKH26 were successfully uptaken by maternal eMSC and pbMSC during a chemotaxis experiment in an agarose spot assay following the procedures of Calle et al [ 11 , 15 ] and Monguió-Tortajada et al [ 16 ] as described in materials and methods ( Figure S5 ).…”

“…Recently, our group characterized for the first time endometrial mesenchymal stem cell lines (eMSC) [ 11 ] as well as peripheral blood MSC (pbMSCs) lines [ 12 ]. While eMSC showed a reduced migratory capacity in response to the implantation cytokine IFN-τ [ 11 ], pbMSC showed chemotactic behavior to both inflammation (TNFα, IL1β), embryo implantation stimuli (IFN-τ) or BBT-secretome, suggesting that the embryo secretome plays a role in ensuring the retention of eMSC and the active recruitment of MSCs from bone marrow during early pregnancy to repress the immune response to prevent the embryo rejection by the maternal organism [ 12 ].…”

“…Therefore, the BBT-9, thanks to its mesenchymal phenotype, could play an immunomodulatory role via EV signaling, aimed at guaranteeing the survival of the embryo. Our group has also reported the immunomodulatory capacity of eMSC and pbMSC by inhibiting the proliferation of human T lymphocytes [ 11 , 12 ].…”

“…Endometrial mesenchymal stem cell (eMSC) lines: eMSC-1A, -3A, -3D, -3E, -4B, -4C, -4D and -4H were isolated, established, and immortalized (PA-317 LXSN-16E6E7 cells) in our laboratory from the uterus of heifers at different oestrous cycle stages [ 11 ].…”

“…The combination of both signals (pro-inflammatory and implantation) would ensure the retention of eMSC in the case of pregnancy. Interestingly, in the absence of embryo stimuli, eMSC showed an apparent mesenchymal to epithelial transition stage [ 11 ]. Additionally, we have just reported the successful isolation for the first time in bovines, of peripheral blood MSC (pbMSCs) lines with classical MSC markers, multipotent capacity, and immune-suppression activity.…”