(756e) Crosslinking of Extracellular Matrix Scaffolds Derived from Pluripotent Stem Cell Aggregates for Neural Differentiation

Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), emerge as promising sources for tissues engineering, drug screening, and disease modeling due to their unique proliferation ability and differentiation potential.  At various developmental stages, PSCs and their progeny secrete large amounts of extracellular matrices (ECM) which could interact with regulatory growth factors to modulate the microenvironment of stem cells.  Hence, ECMs derived from different types of PSC aggregates have broad signaling capacities and can be used as scaffolds to mediate cellular differentiation.  However, the fast degradation of PSC-derived ECMs impedes their wide applications for in vitro cell expansion and in vivo transplantation.  To address this issue, this study investigated the effect of crosslinking on the ECMs derived from PSCs and their regulatory capacity on the proliferation and differentiation of the reseeded PSC-derived neural progenitor cells (NPCs).  To create different biological cues, ECMs from undifferentiated aggregates, spontaneous embryoid bodies, or PSC-derived NPC aggregates were decellularized.  The derived ECM scaffolds were crosslinked using genipin or glutataldehyde to enhance the stability.  The scaffolds were characterized by scanning electron microscopy and atomic force microscopy for topography and elasticity.  PSC-derived NPC aggregates were reseeded on different ECM scaffolds and the cellular compositions of neural progenitors, neurons, and glial cells were characterized.  Human induced pluripotent stem cells were also evaluated for the expressions of pluripotent markers and neural differentiation markers on different ECM scaffolds.  The results indicated that various PSC-derived ECM scaffolds affected stem cell expansion and neural differentiation through intrinsic biological cues and biophysical properties and have potential for in vitro applications and in vivo delivery.

References:

1. Sebastien Sart, Teng Ma, Yan Li.  Extracellular matrices decellularized from embryonic stem cells maintained their structure and signaling specificity. Tissue Eng Part A. 2014, 20: 54-66.

2. Sébastien Sart, Yijun Liu, Teng Ma, Yan Li.  Microenvironment regulation of pluripotent stem cell-derived neural progenitor aggregates by human mesenchymal stem cell secretome. Tissue Eng Part A 2014, In Press. doi:10.1089/ten.TEA.2013.0437.