The General Hospital Corporation

University of Colorado

Mark W Tibbitt

Kristi S Anseth

April M Kloxin

Mehmet Toner

Ajay Shah

Andrea M Kasko

John Oakey

Balaji Sridhar

Disclosed herein are photodegradable hydrogels and associated kits for selectively capturing and releasing cells. The hydrogels result from cross linking in the presence of a photoinitiator (1) a macromer having a polymeric backbone structure, a photo labile moiety, and a first linking moiety, and (2) a cell-binding moiety having a second linking moiety. These two components are cross-linked by a polymerization reaction of the linking moieties to form a photodegradable hydrogel incorporating the cell-binding moiety within the hydrogel. Also disclosed are methods of making the hydrogels, and methods of using the hydrogels for selectively capturing and releasing cells and for detecting cells in a fluid. Such methods can be used to detect the presence and quantity of certain rare cell types in a biological fluid.

Selective capture and release of rare mammalian cells using photodegradable hydrogels in a microfluidic platform

Here, we present a photodegradable microparticle system that can be employed to entrap and deliver bioactive proteins to cells during culture. By using a photosensitive delivery system, experimenters can achieve a wide variety of spatiotemporally regulated release profiles with a single microparticle formulation, thereby enabling one to probe many questions as to how protein presentation can be manipulated to regulate cell function. Photodegradable microparticles were synthesized via inverse suspension polymerization with a mean diameter of 22 &#x3bc;m, and degradation was demonstrated upon exposure to several irradiation conditions. The protein-loaded depots were incorporated into cell cultures and release of bioactive protein was quantified during the photodegradation process. This phototriggered release allowed for the delivery of TGF-&#x3b2;1 to stimulate PE25 cells and for the delivery of fluorescently labeled Annexin V to assay apoptotic 3T3 fibroblasts during culture. By incorporating these photoresponsive protein delivery depots into cell culture, new types of experiments are now possible to test hypotheses about how individual or multiple soluble factors might affect cell function when presented in a uniform, temporally varying, or gradient manner.

Photodegradable groups for tunable polymeric materials

Growing community of inventors

Boulder, CO, United States of America

Mark W Tibbitt