Bioactive Decellularized Extracellular Matrix Hydrogel Microspheres Fabricated Using a Temperature-Controlling Microfluidic System br

Hydrogel microspheres have drawn great attention as functional three-dimensional (3D) microcarriers for cellattachment and growth, which have shown great potential in cell-based therapies and biomedical research. Hydrogels derived from adecellularized extracellular matrix (dECM) retain the intrinsicphysical and biological cues from the native tissues, which oftenexhibit high bioactivity and tissue-specificity in promoting tissueregeneration. Herein, a novel two-stage temperature-controllingmicrofluidic system was developed which enabled production ofpristine dECM hydrogel microspheres in a high-throughputmanner. Porcine decellularized peripheral nerve matrix (pDNM)was used as the model raw dECM material for continuousgeneration of pDNM microgels without additional supportingmaterials or chemical crosslinking. The sizes of the microsphereswere well-controlled by tuning the feed ratios of water/oil phases into the microfluidic device. The resulting pDNM microspheres(pDNM-MSs) were relatively stable, which maintained a spherical shape and a nanofibrous ultrastructure for at least 14 days.Schwann cells and PC12 cells preseeded on the pDNM-MSs not only showed excellent viability and an adhesive property, but alsopromoted cell extension compared to the commercially available gelatin microspheres. Moreover, primary neural stem/progenitorcells attached well to the pDNM-MSs, which further facilitated their proliferation. The successfully fabricated dECM hydrogelmicrospheres provided a highly bioactive microenvironment for 3D cell culture and functionalization, which showed promising potential in versatile biomedical applications