TR1 cells, mesenchymal stem cells and uses thereof
A kind of stem cell and cell technology, applied in the field of Tr1 cells and mesenchymal stem cells, can solve the problem of low degree of proliferation
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Embodiment 1
[0151] Preparation of MSCs
[0152] Mesenchymal stem cells were obtained from the stromal-vascular fraction of adipose tissue of Balb / c mice. Adipose tissue was digested with 0.1% collagenase for 30 minutes and filtered through a 70μ mesh. Adherent cells were cultured in RPMI medium containing 10% FCS and 10% horse serum supplemented with glutamine and penicillin and streptomycin. Cells were frequently monitored for their ability to differentiate into adipocyte and osteoblast lineages.
[0153] Preparation of Tr1 cells
[0154] Ovalbumin-specific Tr1 cells were derived from DO11-10 ovalbumin-specific TCR transgenic mice activated with ovalbumin peptide 323-339 and IL-10 in the presence of irradiated isogenic antigen-presenting cells Differentiate from naive CD4+ T lymphocytes. Cells were then cloned by limiting dilution to obtain a monoclonal population of ovalbumin-specific Trl cells. Cells used in experiments 1 and 2 were obtained from cultures of the Trl clone.
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Embodiment 2
[0162] BALB / c mice were treated with Dextran Sodium Sulfate (DSS, 5% drinking water in water) to induce acute colitis. One group of mice was not treated, or was treated with or without MSCs (0.5×10 6 / mouse) of 10 6 Ovalbumin-specific Tr1 cells were treated intravenously. After 7 days, mice were evaluated for clinical signs based on the following scores:
[0163] 0 - no clinical signs
[0164] 1- Weight Loss
[0165] 2- Weight loss + mild diarrhea
[0166] 3- Weight loss + severe diarrhea
[0167] 4- Weight loss + severe diarrhea + bloody stool
[0168] The results show that co-administration of MSCs can improve the efficacy of Tr1 cell therapy in this model of colitis ( figure 2 ).
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