title: Keratinocyte cytoskeletal roles in cell sheet engineering.
abstract: BACKGROUND
  There is an increasing need to understand cell-cell interactions for cell and tissue engineering purposes, such as optimizing cell sheet constructs, as well as for examining adhesion defect diseases. For cell-sheet engineering, one major obstacle to sheet function is that cell sheets in suspension are fragile and, over time, will contract. While the role of the cytoskeleton in maintaining the structure and adhesion of cells cultured on a rigid substrate is well-characterized, a systematic examination of the role played by different components of the cytoskeleton in regulating cell sheet contraction and cohesion in the absence of a substrate has been lacking.
  
  
  RESULTS
  In this study, keratinocytes were cultured until confluent and cell sheets were generated using dispase to remove the influence of the substrate. The effects of disrupting actin, microtubules or intermediate filaments on cell-cell interactions were assessed by measuring cell sheet cohesion and contraction. Keratin intermediate filament disruption caused comparable effects on cell sheet cohesion and contraction, when compared to actin or microtubule disruption. Interfering with actomyosin contraction demonstrated that interfering with cell contraction can also diminish cell cohesion.
  
  
  CONCLUSIONS
  All components of the cytoskeleton are involved in maintaining cell sheet cohesion and contraction, although not to the same extent. These findings demonstrate that substrate-free cell sheet biomechanical properties are dependent on the integrity of the cytoskeleton network.
authors: Department of Biomedical Engineering, Columbia University, 351 Engineering Terrace, 500 W 120th Street, MC 8904, New York, NY 10027, USA.
authors: Qi
authors: Q
authors: Wei
authors: Daniel
authors: D
authors: Reidler
authors: Min Ye
authors: MY
authors: Shen
authors: Hayden
authors: H
authors: Huang
mesh_terms: D000199:Actins / Q000378:metabolism; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism*; D006801:Humans; D053547:Keratin-14 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D053555:Keratin-5 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D023822:Tissue Engineering
chemical_list: D000199:Actins; D053547:Keratin-14; D053555:Keratin-5; D034741:RNA, Small Interfering