(112b) Direct Production of Human Cardiac Tissues By Pluripotent Stem Cell Encapsulation in Gelatin Methacryloyl
GelMA was synthesized by reacting methacrylic anhydride (MA; 5% w/v) and gelatin for 2 hours, followed by dialysis and freeze-drying. Lyophilized GelMA was re-dissolved into PBS (15% w/v) and combined photoinitiators to form a GelMA precursor solution. Dissociated hiPSCs were combined with the GelMA precursor solution, pipetted into a circular polydimethylsiloxane (PDMS) mold, and photocrosslinked using visible light for 40 seconds, forming circular 3D tissues. Encapsulated hiPSCs were maintained in their pluripotent state for three days followed by the initiation of cardiac differentiation.
Overall, it was seen that the synthesized GelMA successfully photocrosslinked in the presence of hiPSCs and could be used to create 3D cardiac tissues. The first isolated areas of contractions by in the tissues were demonstrated by day 8 of differentiation and these tissues resulted in uniformly contracting tissues by day 20. Confocal microscopy of dissociated tissues showed the presence of large nuclei and well-defined sarcomeres that are indicative of CMs. GelMA is a suitable biomaterial for hiPSC encapsulation and 3D cardiac differentiation. Future work will make use of these cardiac tissues in order to investigate the impacts of drugs on developing CMs in an ontogenic model. This model will allow for in vitro study of a microenvironment very similar to the developing heart.
The results show that GelMA is able to form cell-laden tissues while still maintaining its bioactivity. Using GelMA for both the encapsulation and differentiation, we are able to have only a single cellâ??handling step while still creating functional CMs. As this method requires minimal cell disturbance, it is beneficial for the development and function of 3D human cardiac tissue.
This paper has an Extended Abstract file available; you must purchase the conference proceedings to access it.
Do you already own this?
Log In for instructions on accessing this content.
|AIChE Graduate Student Members||Free|
|AIChE Undergraduate Student Members||Free|