(352e) Embryonic Stem Cell Attachment On Synthetic Chemical Surfaces

Embryonic stem (ES) cells hold great potential in the field of tissue regeneration, cell therapy, and drug discovery due to their self-renewal capacity and differentiation potential. To fully realize the therapeutic potential of ES cells, strategies are needed to maintain the pluripotency of ES cells during expansion. Cell attachment is essential for maintaining ES cells in an undifferentiated state and preventing ES cells from forming unwanted embryoid bodies. Stem cell pluripotency is governed by a unique local microenvironment. ES cells interact with their microenvironment at nanoscale level in vivo . The use of polymers as scaffolds has been well documented and offers several advantages for supporting ES cell maintenance. However, little is known about how surface chemistry influences stem cell pluripotency. In this work, a high-throughput screening platform has been established to study ES cell attachment on a library of synthetic chemical surfaces (66 synthetic chemical surfaces). Using our patented UV graft-induced polymerization method with high throughput (HT), we have synthesized hundreds of new surfaces in days rather than months without the HT. Mouse ES cells were stained with Cell-Tracker Green and seeded into 96-well plates containing the library of 66 synthetic chemical surfaces. The cell attachment on the synthetic surfaces was detected by a Tecan fluorescence microplate reader and normalized with the fluorescence unit of the control group. The surface chemistry with the highest attachment was amines and the lowest attachment was PEGs. The cell attachment on selected membrane was further confirmed by cell morphology observation using optical microscopy and scanning electron microscopy (SEM) as well as cytoskeleton staining. The pluripotency of ES cells grown on the representative membrane was confirmed by retaining pluripotency markers of stem cells, Oct4 and stage-specific embryonic antigen (SSEA-1), which is revealed by immunocytochemistry. Atomic force microscopy and attenuated total reflection infrared spectroscopy were used to examine the topography and degree of grafting of the synthetic chemical surfaces, respectively.