Magnetic cd4 cd62l t cell isolation kit

Chronic colitis was induced by the adoptive transfer of splenic CD4⁺CD25⁻CD62L⁺ T cells into immunocompromised recipient mice as previously described (Heylen et al., 2013 (link)). Previously published work from our research group (Breugelmans et al., 2020 (link)) demonstrated that both the DSS colitis mouse model as well as the adoptive T cell transfer mouse model are adequate models to study intestinal inflammation and barrier dysfunction. For these experiments, we opted for the adoptive T cell transfer model, which is of main interest when studying immunological mechanisms of intestinal inflammation mediated by T cells. Moreover, our research group previously demonstrated the therapeutic potential of helminth-derived products in this experimental animal model of colitis (Heylen et al., 2014 (link); Heylen et al., 2015 (link)). In brief, a single cell suspension was prepared from the spleens of donor BALB/c mice, from which CD4⁺CD25⁻CD62L⁺ T cells were isolated using a magnetic CD4⁺CD62L⁺ T cell isolation kit (Miltenyi Biotec GmhB). To induce colitis, SCID mice were injected i.p. with 1 × 106 CD4⁺CD25⁻CD62L⁺ T cells in 100 µl phosphate-buffered saline (PBS). Control BALB/c mice were i.p. injected with PBS and remained healthy.

Colitis was induced in immunocompromised SCID mice by the adoptive transfer of CD4⁺CD25⁻CD62L⁺ T cells as described previously [25] (link). Briefly, CD4⁺CD25⁻CD62L⁺ T cells were isolated from the spleens of BALB/c donor mice using a magnetic CD4⁺CD62L⁺ T cell isolation kit (Miltenyi Biotec GmbH). To induce colitis, 1×10⁶ CD4⁺CD25⁻CD62L⁺ T cells in 100 µl phosphate-buffered saline (PBS) were intraperitoneally (i.p.) transferred into SCID mice. Two to three weeks after this adoptive transfer, SCID mice started to develop colitis. Control mice were i.p. injected with 100 µl PBS and did not develop colitis [25] (link).