Technical Reference #1949

1949.

Identification of dystroglycan binding sequences in the laminin 2 chain LG4–5 module Nobuharu Suzuki; Kentaro Hozumi; Shunsuke Urushibata; Takashi Yoshimura; Yamato Kikkawa; Jessica D. Gumerson; Daniel E. Michele; Matthew P. Hoffman; Yoshihiko Yamada; Motoyoshi Nomizu, Tokyo University of Pharmacy and Life Sciences, Matrix Biology, 29(1949), (2010)

Keywords:
Laminin; Synthetic peptide; Basement membrane; Heparin; alpha-Dystroglycan

Materials & Methods:
SMG rudiments dissected from embryonic day 12.5 ICR mice were cultured on polycarbonate track-etch filters (13 mm 0.1-μm pore size Sterlitech Corp.) at the air/mediuminterface (Hoffman et al. 2001). The filters were floated on 240 μl of DMEM/F-12 in 50-mm glass-bottom microwell dishes (MatTek). The medium was supplemented with 100 units/ml penicillin 100 μg/ml streptomycin 150 μg/ml vitamin C and 50 μg/ml transferrin. Gland rudiments were cultured on each filter in the presence of 100–150 μg/ml of peptides at 37 °C in a humidified 5% CO2 95% air atmosphere. Glandswere photographed after 2 h and 40 h and the number of end buds was counted and expressed as a ratio (the number of end buds at 40 h/number at 2 h).

Cell Type(s)
Ex vivo SMG organ culture