Figure 2. Mononuclear cell numbers after enzymatic dissociation methods in mice with clinical experimental autoimmune encephalomyelitis.

(A) All enzymatic dissociation methods yielded high enough total live cell numbers to meet our second criterion, and accutase 15/papain 30 showed significantly higher numbers of live cells than accutase 15, papain 15, papain 30 and papain 60. After enumeration of total live cells, Kit treatments yielded a higher median cell number than accutase 15/papain 30. (B) Treatment of spinal cord tissue of mice with clinical experimental autoimmune encephalomyelitis with different dissociation methods demonstrated that the Kit and papain 60 yielded sufficient cell numbers for further flow cytometiic analyses as pre-defined by the second criterion. (C) When mononuclear cells isolated from the brain were gaited on CD45⁺ cells, Kit enzymatic dissociation yielded a higher median cell number than other methods, including accutase 15/papain 30. (D) In the spinal cord, numbers of CD45⁺ cells were similar between Kit and papain 60. Groups were compared for normality by using the Kolmogorov-Smirnov test. Values were compared using the non-parametric test Mann-Whitney U test. *p < 0.05, **p < 0.01, ***p < 0.001.