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. Author manuscript; available in PMC: 2013 Oct 31.
Published in final edited form as: J Immunol Methods. 2012 Jul 16;384(1-2):157–163. doi: 10.1016/j.jim.2012.07.002

Figure 1. Comparison of strategies to extract and enrich gastric leukocytes.

Figure 1

Gastric tissue from H. pylori infected mice were subjected to mechanical processing, Dispase II (Disp.), Collagenase IV (Coll. IV) or Collagenase IV plus EDTA/DTT digestion (EDTA/DTT/Coll. IV). Mechanically dissociated tissue and collagenase IV digested tissue released the greatest number of total cells, while dispase treatment resulted in the fewest (A). The total number of CD45+ cells was highest after collagenase IV and collagenase IV/EDTA/DTT treatment (B). CD4+ cell numbers are significantly higher with collagenase IV and collagenase IV/EDTA/DTT treatment (C). CD8+ cell number was greatly decreased with dispase digestion while collagenase IV with EDTA/DTT digestion yielded the greatest amount of CD8+ cells (D). Lymphocyte enrichment was performed using a discontinuous Percoll or lympholyte M gradient (E). More cells were obtained using lympholyte M enrichment and greater percentages of CD45+ and CD3+ cells were also obtained using this gradient. Graphs shown represent 3 mice per group and are presented as mean ± standard error. Groups were compared using one-way analysis of variance (ANOVA) followed post-hoc by Tukey multiple comparison tests on log-transformed data. Significant differences are represented by the following *p<0.05, **p<0.01, ***p<0.001. Data shown are representative of 3 independent experiments performed on 3 occasions in total.