Collagen type 1 primary antibody

After 4 d of culture on titanium discs, hDPSCs were washed with PBS and fixed in 4 % paraformaldehyde (PFA) for 10 min at room temperature. The cells were then incubated with 10 % goat serum (Invitrogen, CA, USA) for 10 min at room temperature, followed by overnight incubation at 4 °C with either Ki67 primary antibody (Rabbit polyclonal, Abcam, Cambridge, UK) or collagen type I primary antibody (Rabbit polyclonal, Abcam, Cambridge, UK) at 1:500 dilution using 0.1 % Triton X-100/1 %BSA/PBS. The secondary antibody, alexa fluor 488 conjugated rabbit IgG (Abcam, Cambridge, UK), was used for primary antibody localisation for 1 h at room temperature. Nuclei were stained with 4',6-diamino-2-fenilindol (DAPI, Invitrogen, CA, USA). The images were taken using an Axioskop fluorescence microscope (Zeiss, Overkochen, Germany) and Nikon DS-Qi1 camera (Nikon, Tokyo, Japan).