Computational protocol: Retention and Functional Effect of Adipose-Derived Stromal Cells Administered in Alginate Hydrogel in a Rat Model of Acute Myocardial Infarction

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Protocol publication

[…] Immediately following animal sacrifice by decapitation during anesthesia, the heart was excised, washed in saline, retrograde perfusion from the aorta was performed with saline followed by 4% paraformaldehyde. The heart was then placed in formaldehyde for 24 hours and stored in 70% ethanol before embedding in paraffin. The hearts were cut in the short axis on a microtome, and immunohistochemistry was performed using the following: CD31 (Novus Bio, NB100-2284), CD68 (Abcam, 125212), Masson's trichrome, and hematoxylin and eosin (H&E). Prior to antibody staining, antigen retrieval was performed using citrate buffer and microwave treatment. Images were obtained by an ×20 objective scanning and subsequent stitching, producing images of the entire short axis slice. Fibrosis on Masson's trichrome stainings was quantified using ImageJ (Fiji) on the entire heart. CD31 clusters in peri-infarct regions were scored from 1–4, and CD68 was counted in the whole infarct area and normalized to the counted area. [...] For functional measurements, changes in measured parameters within groups were assessed using paired t-test with Bonferroni correction, while comparisons between groups were performed using univariate analysis with day 1 as covariate and Tukey as post hoc test. Retention was compared using repeated measures ANOVA with Tukey post hoc test. All statistics were performed using SPSS (IBM SPSS statistics v. 22). […]

Pipeline specifications

Software tools Stitching, ImageJ, SPSS
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Rattus norvegicus, Homo sapiens
Diseases Heart Diseases, Myocardial Infarction