Takagi Ryo    Department   Research Institutes and Facilities, Research Institutes and Facilities    Position   Assistant Professor
Language	Japanese
Title	Laminin-521 maintains proliferation activity of cultured human stratified squamous epithelial cells with less effect of individual difference.
Conference	The 48th annual meeting of the Japanese society of matrix biology and medicine
Conference Type	Nationwide Conferences
Presentation Type	Speech
Lecture Type	General
Date	2016/06/24
Summary	[Aim] Cultured stratified squamous epidermal cells are known as useful ectopical and/or allogeneic cell graft in clinical study. Therefore, development of culture method for proliferation of the epithelial cells is important in the area of regenerative medicine. In the present study, human type I collagen (placenta), human type IV collagen (placenta), and recombinant human laminin-521 were used for colony formation, cell adhesion, and proliferation assay of human stratified epithelial cell. [Methods] The ECM proteins were coated on culture dishes at 1 µg/cm2 over night at 4ºC. Normal culture dishes were used as control. Stratified squamous epithelial cells were prepared from human oral mucosal and skin tissues and cultured in low [Ca2+] medium (KGM-Gold). NIH-3T3 treated with mitomycin C was used as a feeder layer for primary culture of the epidermal keratinocytes. [Results] Primary human keratinocytes showed significantly higher colony forming efficiency on these ECM coated dishes than control. Although there is no significant difference in adhesion activities of the cells cultured over night among control and ECMs, the activities in cultivation for 1h were significantly promoted by ECMs. In the results of subculture, proliferation activity of the epithelial cells cultured on laminin-521 dish was significantly higher than control, and laminin-521 maintained proliferation activity of the epithelial cells with less effect related to individual variability among the human tissues in comparison to type I and IV collagen. [Conclusion] Laminin-521 is indicated as one of a candidate factor for maintaining proliferation activity of cultured human stratified squamous epithelial cells.