RESUMO
We cultured isolated islets from human or porcine origin in the presence or absence of IL1 and TNFα and studied cytoprotective effects of two structurally different PBR ligands. Storage of pig or human islets in the presence of cytokines significantly lowered the fraction of vital beta-cells. Compared with cytokine incubations PK11195 alone or in combination with cytokines was effective to prevent cytokine induced cell death. The data indicate that cold storage in the presence of PK11195 may further protect beta-cells from cytokine induced cell death. This ligand may be helpful to preserve beta-cell survival before transplantation.
Assuntos
Citocinas/farmacologia , Citoproteção/efeitos dos fármacos , Células Secretoras de Insulina/citologia , Células Secretoras de Insulina/efeitos dos fármacos , Isoquinolinas/metabolismo , Isoquinolinas/farmacologia , Receptores de GABA-A/metabolismo , Animais , Morte Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Temperatura Baixa , Citometria de Fluxo , Humanos , Células Secretoras de Insulina/transplante , Transplante das Ilhotas Pancreáticas , Ligantes , Especificidade por Substrato , Suínos , Preservação de TecidoRESUMO
Incompatible blood group antigens are highly immunogenic and can cause graft rejections. Focusing on distinct carbohydrate- and protein-based membrane structures, defined by blood group antigens, we investigated human bone marrow-derived mesenchymal stem cells (MSCs) cultured in human serum. The presence of H (CD173), ABO, RhD, RhCE, RhAG, Kell, urea transporter type B (SLC14A1, previously known as JK), and Duffy antigen receptor of chemokines (DARC) was evaluated at the levels of genome, transcriptome and antigen. Fucosyltransferase-1 (FUT1), RHCE, KEL, SLC14A1 (JK) and DARC mRNA were transcribed in MSCs. FUT1 mRNA transcription was lost during differentiation. The mRNA transcription of SLC14A1 (JK) decreased during chondrogenic differentiation, while that of DARC increased during adipogenic differentiation. All MSCs synthesized SLC14A1 (JK) but no DARC protein. However, none of the protein antigens tested occurred on the surface, indicating a lack of associated protein function in the membrane. As A and B antigens are neither expressed nor adsorbed, concerns of ABO compatibility with human serum supplements during culture are alleviated. The H antigen expression by GD2dim+ MSCs identified two distinct MSC subpopulations and enabled their isolation. We hypothesize that GD2(dim+) H(+) MSCs retain a better 'stemness'. Because immunogenic blood group antigens are lacking, they cannot affect MSC engraftment in vivo, which is promising for clinical applications.