Hollow Hydrogel Cocoons for the Encapsulation of Therapeutic Cells Using a Microfluidic Platform

Abstract

Microencapsulation of stem cells in hydrogel for use in therapeutic applications has been shown to improve cell retention at the site of injuries due to their mechanical and immunoprotective properties. These microscale droplets (cocoons) can be produced at high throughputs within microfluidic channels. Currently, the ability for cells to egress hydrogel cocoons is under investigation. This egress can correlate with therapeutic efficacy, and so promoting or inhibiting the egress of cells can be a vital component of viable treatments. Previously, a second hydrogel layer was shown to reduce egress, but issues involving cell proliferation were unchanged. We propose a microfluidic process to encapsulate cells in two layers of thermoresponsive hydrogels, in which the inner core melts at physiological temperatures to form hollow cocoons that allow cells free motion inside the immunoprotective shell. We hypothesize that the open volume would increase cell viability and proliferation, without increasing cell egress due to the uninterrupted hydrogel shell. In this project the encapsulation of NIH 3T3 cells in hollow agarose cocoons was achieved. 3T3 cells were first encapsulated in thermoreversible gelatin which were then re-encapsulated in agarose through the use of a flow-focusing microfluidic channel with on-chip mixing of two inlet flows to produce hollow cocoons. The production of these cocoons showed the potential of high throughput, monodisperse samples with future investment. Preliminary investigation in the behavior of the encapsulated cells showed that the cells maintain high viability over the course of 48 hours. There are early indications that the hollow nature of correctly formed cocoons can limit cell egress, and may allow for proliferation in the cocoon.