Protein functionalization and 3 D cell culturing with

Protein functionalization and 3 D cell culturing with PEG Ilyssa Evans Spring 2016

Background Polyacrylamide for Cell Culturing Node Pore Sensing u u Low-cost method that measures transit times of cell as they interact with different antibodies throughout a channel Can be used to determine the phenotypes of cells Goal: functionalize and pattern proteins/antibodies in a microfluidic channel using UV light Stiffness Factor u u “Stiffness” of tissue in our body naturally varies Varying stiffness of extracellular matrix has great impact on traits of anchorage -dependent cells (MCF 7) Goal: find a suitable hydrogel for cells to grow on that can easily have it’s stiffness changed

Refresher

Methods – Part I Functionalization of PDMS Devices Photoinitiator Tape + UV Light m. Cherry (with –SH group) X 4 PBS+ Fluorescence

Is it Traut’s reagent? Beginning of Channel Non-Trauts Middle of Channel Non-Trauts

Is functionalization happening? Multiple Iterations Plasma Silane Protein 15 minutes 24 hours Silane Plasma Protein Silane Protein

New Approach to Cell Culturing PEG!

PEG Hydrogels General u u Polyethylene glycol is a polyether with many biomedical applications Water-soluble Polyethylene glycol monoacrylate Polyethylene glycol diacrylate PEG!

Methods – Part II Fabrication of PEG Hydrogels

Methods – Part II Fabrication of PEG Hydrogels + Cell Encapsulation

Cell Encapsulation PEG Cell Encapsulation – 72 hours Trial 1 Trial 2 Trial 3

Conclusion Polyacrylamide for Cell Culturing Protein Conjugation u No issues with the protein conjugation appears to be happening – it should work u Retry the original method – acrylating the channel instead of the glass or PDMS beforehand. u Need to try with photoinitiator for patterning PEG Hydrogels u Hydrogel fabrication was a success u Need to repeat 3 D cell patterning and attempt 2 D cell patterning

Methods – Part I Functionalization of PDMS Devices