On-line observation of cell growth in a three-dimensional matrix on surface-modified microelectrode arrays.
Despite many successful applications of microelectrode arrays (MEAs), typical two-dimensional in-vitro cultures do not project the full scale of the cell growth environment in the three-dimensional (3D) in-vivo setting. This study aims to on-line monitor in-vitro cell growth in a 3D matrix on the surface-modified MEAs with a dynamic perfusion culture system. A 3D matrix consisting of poly(ethylene glycol) hydrogel supplemented with poly-D-lysine was subsequently synthesized in situ on the self-assembled monolayer modified MEAs. FTIR spectrum analysis revealed a peak at 2100 cm(-1) due to the degradation of the structure of the 3D matrix. After 2 wks, microscopic examination revealed that the non-degraded area was around 1500 microm(2) and provided enough space for cell growth. Fluorescence microscopy revealed that the degraded 3D matrix was non-cytotoxic allowing the growth of NIH3T3 fibroblasts and cortical neurons in vitro. Time-course changes of total impedance including resistance and reactance were recorded for 8 days to evaluate the cell growth in the 3D matrix on the MEA. A consistent trend reflecting changes of reactance and total impedance was observed. These in-vitro assays demonstrate that our 3D matrix can construct a biomimetic system for cell growth and analysis of cell surface interactions.
Lin
Shu-Ping
SP
Institute of Biomedical Engineering, National Cheng Kung University, 1 Ta-Hsueh Rd., Tainan 701, Taiwan, ROC.
Kyriakides
Themis R
TR
Chen
Jia-Jin J
JJ
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D032701:Biomimetics; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D017097:Electric Impedance; D004867:Equipment Design; D005347:Fibroblasts / Q000378:metabolism; D005456:Fluorescent Dyes / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D007211:Indoles / Q000378:metabolism; D051379:Mice; D008839:Microelectrodes; D008856:Microscopy, Fluorescence; D041681:NIH 3T3 Cells; D009474:Neurons / Q000378:metabolism; D009862:Online Systems; D010477:Perfusion; D010590:Phalloidine / Q000378:metabolism; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000737:chemistry; D012235:Rhodamines / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared; D013379:Substrate Specificity; D013499:Surface Properties; D013997:Time Factors
D001672:Biocompatible Materials; D005456:Fluorescent Dyes; D007211:Indoles; D012235:Rhodamines; D010590:Phalloidine; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; D011092:Polyethylene Glycols; C007293:DAPI; D006046:Gold