title: Retrieval of rat aortic smooth muscle cells as intact cell sheet for regenerative medicine: a cost effective approach using photo polymerization.
abstract: Cell-based therapeutics are promising routes for the regeneration of damaged cells and organs. The recovery of cells cultured in vitro for such applications requires the use of proteolytic enzymes which deteriorate its property by disruption of cell-cell and cell-matrix interactions. Intact cell sheets can be retrieved with the use of thermo responsive polymer grafted on to the culture plates. Our study presents the use of photo-polymerization as a simple and inexpensive way to create thermo-responsive culture surfaces for the detachment of intact cell sheet. Poly (N-isopropyl acrylamide) (PNIPAAm) was synthesized by photo-polymerization and characterized by NMR spectroscopy, differential scanning calorimetry and gel permeation chromatography. Thermo-responsive culture dishes were prepared by the coating method and characterized for its thermo-responsive efficacy using FTIR spectroscopy and water contact angle measurements. Atomic force microscopy depicted the thin coating achieved with this method is similar to the conventional grafting method. Suitability for cell culture and cell sheet retrieval was assessed by culturing rat aortic smooth muscle cells in the PNIPAAm coated tissue culture plates. The cells remained viable as evident from the live dead assay and the cell sheet was detached by low temperature treatment. The results demonstrate a versatile method for creating thermo responsive culture surfaces while eliminating the use of expensive radiation sources for the conventional grafting method.
authors.affiliation: Division of Tissue Engineering and Regeneration Technologies, Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram, Kerala 695012, India.
authors.forename: Lakshmi M
authors.initials: LM
authors.lastname: Mukundan
authors.forename: Remya
authors.initials: R
authors.lastname: Nirmal
authors.forename: Lynda V
authors.initials: LV
authors.lastname: Thomas
authors.forename: U S
authors.initials: US
authors.lastname: Sajeev
authors.forename: Prabha D
authors.initials: PD
authors.lastname: Nair
mesh_terms: D000178:Acrylamides / Q000737:chemistry* / Q000528:radiation effects; D000180:Acrylic Resins; D000818:Animals; D001011:Aorta / Q000166:cytology*; D002152:Calorimetry, Differential Scanning; D060846:Cell Engineering / Q000379:methods*; D002470:Cell Survival; D002478:Cells, Cultured; D018625:Microscopy, Atomic Force; D032389:Myocytes, Smooth Muscle / Q000166:cytology*; D019906:Nuclear Magnetic Resonance, Biomolecular; D055668:Photochemical Processes; D058105:Polymerization; D011108:Polymers / Q000737:chemistry* / Q000528:radiation effects; D011137:Polystyrenes / Q000737:chemistry; D051381:Rats; D044968:Regenerative Medicine / Q000191:economics / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature
chemical_list: D000178:Acrylamides; D000180:Acrylic Resins; D011108:Polymers; D011137:Polystyrenes; C052970:poly-N-isopropylacrylamide