An Optimized Confined Singie-Cell Patterning in Simple Reversible Microflu-idic Chip
Objective To obtain the confined single-cell patterned multiple cellular arrays exposed to exactly controllable fluidic shear stress stimuli.Methods Based on micro-contact printing(μCP),3D printed mold and vacuum negative bonding,a simple microfluidic chip with single-cell patterned osteoblasts was developed.Meanwhile,μCP-controled cell micro-patterns were optimized and deter-mined exactly,from two main aspects of protein patterning and cell patterning,with altering related key parameters in terms of plasma treatment time,FN coating time,cleaning times,printing time,F127 concentration,F127 treatment time,cell seeding time and cell den-sity.Results The optimized parameters for good FN micro-patterns were plasma oxygen treatment for 1 minute,FN coating for 30 min-utes,stamp contact time of 6-24 hours,and 2 washes.The optimal parameters for confined micro-patterned cellular arrays were 2％F127,F127 incubation for 1 hour,cell density of 2×106 mL-1,and cell seeding for 15-30 minutes.The confined ostoeblastic patterns can maintain for 6-8 hours.The microfluidic chip based on 3D printed molds and vacuum sealing could provide two different fluid shear forces simultaneously.Conclusion This new and developed method provides a promising experimental platform for studying the reaction between patterned multicellular system and mechanical microenvironment.
万方数据
维普
