Local mechanical stimulation of Mardin-Darby canine kidney cell sheets on temperature-responsive hydrogel

• Published in: International journal of molecular sciences Vol. 13; no. 1; pp. 1095 - 1108
• Main Authors: Harada, Ichiro, Yanagisawa, Shunpei, Iwasaki, Katsuhiko, Cho, Chong-Su, Akaike, Toshihiro
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.01.2012; Molecular Diversity Preservation International (MDPI)
• Subjects: Acrylic Resins - chemistry; Animals; Cadherins - metabolism; Cell Adhesion; Cell Line; Cell Movement; Cell Shape; Cells; Collagen; Collagen - chemistry; collective cell migration; Communication; Dogs; E-cadherin; Hydrogel, Polyethylene Glycol Dimethacrylate - chemistry; Hydrogels; MDCK cell sheets; mechanical stimulation; Mitogen-Activated Protein Kinase 1 - metabolism; Mitogen-Activated Protein Kinase 3 - metabolism; Motility; Phase transitions; Phosphorylation; Signal Transduction; Stress, Mechanical; Temperature; temperature-responsive hydrogel; MDCK cell sheets; collective cell migration; mechanical stimulation; temperature-responsive hydrogel; E-cadherin
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms13011095

Collective motion of cell sheets plays a role not only in development and repair, but also in devastating diseases such as cancer. However, unlike single-cell motility, collective motion of cell sheets involves complex cell-cell communication during migration; therefore, its mechanism is largely unknown. To elucidate propagation of signaling transduced by cell-cell interaction, we designed a hydrogel substrate that can cause local mechanical stretching of cell sheets. Poly (N-isopropyl acrylamide) (PNIPAAm) hydrogel is a temperature-responsive polymer gel whose volume changes isotropically in response to temperature changes below 37 °C. We designed a combined hydrogel substrate consisting of collagen-immobilized PNIPAAm as the local stimulation side and polyacrylamide (PAAm) as the non-stimulation side to assess propagation of mechanical transduction. Mardin-Darby canine kidney (MDCK) cells adhered to the collagen-immobilized PNIPAAm gel increased it area and were flattened as the gel swelled with temperature decrease. E-cadherin in these cells became undetectable in some domains, and actin stress fibers were more clearly observed at the cell base. In contrast, E-cadherin in cells adhered to the collagen-immobilized PAAm side was equally stained as that in cells adhered to the collagen-immobilized PAAm side even after temperature decrease. ERK1/2 MAPK activation of cells on the non-stimulated substrate occurred after partial stretching of the cell sheet suggesting the propagation of signaling. These results indicate that a change in the balance of mechanical tension induced by partial stretching of cell sheets leads to activation and propagation of the cell signaling.