Thermo-responsible copolymer interaction with human induced pluripotent stem cells for functional and pyramidal neuron differentiation

6^th International Conference and Exhibition on Materials Science and Engineering

September 12-14, 2016 Atlanta, USA

Kapil Dev, Yuri Rochev, Alexander Gorelov and Sanbing Shen

National University of Ireland, Ireland
University College Dublin, Ireland

Posters & Accepted Abstracts: J Material Sci Eng

Abstract :

The interaction among human induced pluripotent stem cells (hiPSCs) with thermo-responsible copolymer substrates signifies to have a reflective impact on stem cells based applications in regenerative medicine and tissue engineering. Development and design of noval biomaterial that should be non-toxic and having unique properties that could drive stem cell fate both in vivo and in vitro is biggest challange. We report thermo-responsible solvent casting copolymer i.e. poly(N-isopropylacrylamide)(NiPAM)-co-poly(N-tert-butlyacrylamide) (NtBAM) system for guiding the hiPSCs more into pyramidal or cortical neurons fate. A three conditions (Control for iPSCs i.e. Geltrex; copolymer and Geltrex coated copolymer) was designed to carryout whole experiment using three iPSCs donors (1C-C1, 02V-C1 and 3VC1). Presence of Geltrex, Copolymer and Geltrex coated copolymer were charachertized using Fourier transform infrared spectroscopy (FITR) and Scanning Electron Microscope (SEM). FTIR and SEM shows the presence of NIPAM, NtBAM and NIPAM-co-NtBAM on culture vessels. Atomic Force Microscope (AFM) was used to determine the roughness of Geltrex (48.13 nm), copolymer (33.07 nm) and blank (34.58 nm, without copolymer or Geltrex), respectively. Where thickness of Geltrex (338 nm) and copolymer (792.84 nm) were measured by AFM. An experiment for 15 days were designed for hiPSCs behaviour on three conditions. Geltrex cultured hiPSCs were in typical round, colony shaped till day 12 and found positive for pluripotent markers (OCT4, SOX2 and NANOG) with slightly differentiated cells; whereas hiPSCs on copolymer were change their morphological pattern from day one and differentiated into typical pyramidal neurons on day 12 and were positive with neuronal i.e. TUJ1, NESTIN, DCX, MAP2, ASM (Mesoderm) and AFP (Endoderm) markers using immunostaining, Western blot and qt-PCR methods. hiPSCs on Geltrex coated copolymer were in intermediated stage of iPSCs and neuronal differentiation. Hence, we conclude that there is strong correlation among laboratory designed copolymer and iPSCs differentiation into neuronal fate and NIPAM-co-NtBAM enchances the neuronal differentiation process i.e. neuronal stem cells, neuronal precursor cells or terminal differentiated neuronal cells. This specially designed copolymer system may pave a way for future rapid differentiation process and in the treatment of neuronal diseases.

Biography :

Email: kapildchauhan@yahoo.com