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. 2018 May 18;9(38):25034–25047. doi: 10.18632/oncotarget.25244

Figure 4. Illustration of the sponge-gel-supported histoculture method and immunohistochemistry read-out.

Figure 4

(A) Biopsies from previously untreated HNSCC patients are taken under general anaesthesia after informed consent. Biopsies are transported to the laboratory within 1 hour and cut into single fragments. Each single fragment is cultured on a sponge drenched into medium in a 12-well plate. The fragment is placed on the sponge in such a way that it is surrounded by air and attached to the sponge enabling it to absorb medium. Using this method, the in-vivo situation is simulated. (B) Illustration of the pathological scoring system. Of each single tumor fragment, at day 0 and 7, three slides are cut and stained for pan-Cytokeratin, Hematoxylin and Eosin (H&E) and Ki-67. With the H&E and pan-Cytokeratin staining the percentage of tumor is scored (% Tumor, cancer cells). With the H&E staining the percentage of viable cancer cells in relation to the total amount of tumor (including areas of necrosis) is determined (% Viability). The Ki-67 staining is used to determine the proliferation rate of the viable cancer cells (% Proliferation).