Colorectal cancer and primary fibroblast, pericyte conditioned media
Proteomic profiling for 200 human cytokines of conditioned media samples as follows: PM = pericyte-conditioned pericyte media PCM = Caco2-conditioned pericyte media PSM = SW480-conditioned pericyte media PHM = HCT116-conditioned pericyte media FM = fibroblast-conditioned fibroblast media FCM = Caco2-conditioned fibroblast media FSM = SW480-conditioned fibroblast media FHM = HCT116-conditioned fibroblast media
Steps to reproduce
Proteomics data was generated thanks to RayBiotech quantitative proteomics services, QAH-CAA-4000. Base medias used for this study are complete pericyte media (ScienCell Cat#1201) and complete fibroblast media (Lonza Cat#CC-3132). Cells used for this study are Caco-2 (ATCC HTB-37), SW480 (ATCC CCL-228), HCT-116 (ATCC CCL-247), human brain vascular pericytes (ScienCell Cat#1200), and normal human lung fibroblasts (Lonza CC-2512) Conditioned Media Generation and Proteomics: For all cell types, 1.5e6 cells were plated onto a 115 cm2 tissue culture dish with 15 mL of their respective media and allowed to attach for 24 hours. After this time, media was removed and replaced with either fibroblast or pericyte complete growth medium to generate conditioned fibroblast media and conditioned pericyte media. This conditioned media was gathered after 24 hours to balance adequate cytokine content and nutrient depletion. Conditioned media were centrifuged for ten minutes at 800 rcf to remove dead cells and large cell debris, and supernatants were aliquoted and frozen at -80° C until use. Quantitative proteomic profiling was performed using testing services from RayBiotech (200 Human Proteins Q4000, RayBiotech, Inc., Norcross, GA).