Hepato stim medium

Patients undergoing scheduled surgery at Christiana Care Health System (Newark, DE) consented for an unaffected portion of their parotid gland tissue to be transferred to Rice University or the University of Texas Health Science Center at Houston under IRB-approved protocols. The fresh parotid gland tissue was prepared in agreement with a standard operating protocol for generating hS/PCs (Wu et al., 2018 (link)). hS/PCs were cultured in Hepato-STIM™ medium supplemented with 10 ng/mL EGF (355056; Corning) and 1% (v/v) penicillin-streptomycin (15140122; Life Technologies/ThermoFisher), and maintained at 37°C in a 5% (v/v) CO₂ incubator as described previously (Pradhan et al., 2009 (link)). The studies in this article used samples from three female donors age, 22, 57 and 63. hS/PCs expressed biomarkers, K5, K14, and p63 and were encapsulated in hydrogels and cultured in complete Hepato-STIM™ medium for these studies. A full characterization of these cells appeared in Srinivasan et al. (2017 (link)) and they were fully sequenced in the functional annotation of the mammalian genome 5 (FANTOM5) project (FANTOM Consortium the RIKEN PMI CLST et al., 2014 (link)).

Human salivary gland tissues were procured from consented patients undergoing surgery for head and neck cancer following protocols approved by institutional review boards at both Christiana Care Health Systems and the University of Delaware. Explant culture for the isolation of hS/PCs was adapted from our previous isolation protocols 29, 31. Human parotid gland (hPG) tissue was disinfected with 1% (vol/vol) betadine solution in cold Dulbecco's Modified Eagle Medium (DMEM)/Nutrient Mixture F‐12 (Thermo Fisher Scientific, Grand Island, NY,
https://www.thermofisher.com), minced into a slurry of small pieces and suspended in hepato‐STIM medium (Corning Inc. ‐ Life Sciences, Oneonta, NY,
http://www.corning.com) supplemented with 1% (wt/vol) penicillin‐streptomycin, 1% (vol/vol) Fungizone (Thermo Fisher), and epidermal growth factor (EGF) (10 ng/ml). On reaching 70%–80% confluence, cells were trypsinized using 0.05% (wt/vol) trypsin‐EDTA (Thermo Fisher) that was stopped by adding trypsin soybean inhibitor (Sigma‐Aldrich, St‐Louis, MO,
http://www.sigmaaldrich.com). Passages 2 through 15 were used for this study and all the experiments were repeated with cells isolated from at least 3 different patients.