Tumor Tissue Collection and Culture

Tumors were collected following informed patient consent at Thomas Jefferson University Hospital under an IRB-approved protocol (#07D.483). The following PTC samples were isolated: TJU-THY #1 is from a 53 y/o, TJU-THY#2 is from a 32 y/o female, and TJU-THY#3 is from a 38 y/o. Less than 16 h post-surgery, excess adipose and stromal tissue was removed and tumors were cut into 1 mm³ pieces. Vetspon absorbable hemostatic gelatin 1 cm³ sponges (Novartis; Basel, Switzerland) were pre-soaked in 12-welled plates for 15 min at 37 °C in 500 μL of DMEM/10% FBS containing drugs or DMSO as a vehicle control. To avoid concerns of intratumoral heterogeneity, up to three 1 mm³ pieces from different locations of the original tumor were placed per sponge per treatment condition. Medium was replaced every 24 h. Tumor pieces for western blotting were homogenized in modified RPPA lysis buffer [75 (link)] with phosphatase and inhibitors (PhosSTOP and cOmplete tablets Roche, Basel, Switzerland). Laemmli sample buffer was added, and samples were heated at ≥95 °C for 5 min. In parallel, the lysates were sent to MD Anderson and used for reverse phase protein array (RPPA) analysis as previously described [46 (link)]. Data was visualized using Morpheus (Broad Institute).