Adipose-derived mesenchymal stromal cells from genetically modified pigs: immunogenicity and immune modulatory properties

作者：Kumar Goutham; Hara Hidetaka; Long Cassandra; Shaikh Humza; Ayares David; Cooper David K C; Ezzelarab Mohamed^*

来源：Cytotherapy, 2012, 14(4): 494-504.

DOI：10.3109/14653249.2011.651529

• alpha 1,3-galactosyl transferase knock-out
• adipose-derived
• mesenchymal stromal cells
• pig
• xenotransplantation

Background aims. The immunomodulatory and anti-inflammatory effects of mesenchymal stromal cells (MSC) could prove to be a potential therapeutic approach for prolongation of survival of cell xenotransplantation. Adipose (Ad) MSC from genetically modified pigs could be an abundant source of pig donor-specific MSC. Methods. Pig (p) MSC were isolated from adipose tissue of a 1,3-galactosyltransferase gene knock-out pigs transgenic for human (h) CD46 (GTKO/hCD46), a potential source of islets. After characterization with differentiation and flow cytometry (FCM), AdMSC were compared with bone marrow (BM) MSC of the same pig and human adipose-derived (hAd) MSC. The modulation of human peripheral blood mononuclear cell (hPBMC) responses to GTKO pig aortic endothelial cells (pAEC) by different MSC was compared by measuring 3H-thymidine uptake. The supernatants from the AdMSC cultures were used to determine the role of soluble factors. Results. GTKO/hCD46 pAdMSC (i) did not express galactose-alpha 1,3-galactose (Gal) but expressed hCD46, (ii) differentiated into chondroblasts, osteocytes and adipocytes, (iii) expressed stem cell markers, (iv) expressed lower levels of Swine Leucocyte Antigen I (SLAI), Swine Leucocyte Antigen II DR (SLAIIDR) and CD80 than pAEC before and after pig interferon (IFN)-gamma stimulation. The proliferative responses of hPBMC to GTKO/hCD46 pAdMSC and hAdMSC stimulators were similar, and both were significantly lower than to GTKO pAEC (P < 0.05). The proliferation of hPBMC to GTKO pAEC was equally suppressed by GTKO/hCD46 pAdMSC and hAdMSC (P < 0.05). The supernatant from GTKO/hCD46 pAdMSC did not suppress the human xenoresponse to GTKO pAEC, which was cell-cell contact-dependent. Conclusions. Initial evidence suggests that genetically modified pAdMSC function across the xenogeneic barrier and may have a role in cellular xenotransplantation.

• 出版日期2012-4