Figure 3

3D culture and in vivo results on tumor viability using secretome formulation. (a) MTT results of dose dependent effect of SF (freeze dried secretome in Mammocult control) on TNBC spheroids after 48 h showing MIC value above 30 mg/ml. (b) Graphical plot of tumor volumes of each animals in three groups (control Ringer’s lactate saline, dose1-50 mg/ml and dose2-100 mg/ml); n = 6 for each group. Intratumoral inection was given at tumor size of 1 mm3 and further tumor growth was absent only when SF was injected. (c) Results of flow cytometry to analyse CD44⁺/CD24⁻ breast cancer stem cells, multi drug resistance protein 1 (MDR1), programmed death ligand-1(PD-L1) expressing cells in xenograft tissue. Drastic decrease in all markers was found in treated cancer tissue. (d) Representative images of hematoxylin and eosin stained tumor from each group showing blue stained tumor necrotic regions (see arrows pointing to these regions) in the treated cancer tissue samples. Surrounding the necrotic regions are regions cleared of cancer tissue and populated by murine cells. The pink stained tumor stroma is not visible in these regions. In the control, that is saline injected, the arrow points to cancer tissue with extensive pink-stained cancer stroma. The stroma is filled with cancer cells. All graphical plots are mean ± SD values and differences between control and treatment concentrations were considered statistically significant for p value < 0.05 and is represented by (*).