MSCs were seeded at 1 × 105 cells/well on the fabricated pGelMA hydrogel pyramid model after
ASP at 10 s, 5 c in a 24-well plate, and cultured at 37 °C under
a humidified 5% CO2 atmosphere. After culturing, the culture
medium was removed, and the hydrogels were washed with D-PBS and fixed
with 4% paraformaldehyde. After washing with water, the hydrogel model
was permeabilized with 0.5% Triton X-100. After being washed with
D-PBS, the specimens were stained with 100 nM rhodamine-phalloidin
(Fujifilm Wako Pure Chemical Industries, Ltd., Osaka, Japan) for 40
min in the dark. After washing 3 times with D-PBS, the hydrogels were
stained with 4′,6-diamidino-2-phenylindole, dihydrochloride
(Dojindo Laboratories, Kumamoto, Japan) diluted 1:500 in D-PBS for
5 min in the dark. After washing with D-PBS, the hydrogels were observed
by using a confocal laser scanning microscope (CLSM) (FV3000, EVIDENT,
Tokyo, Japan).
ASP at 10 s, 5 c in a 24-well plate, and cultured at 37 °C under
a humidified 5% CO2 atmosphere. After culturing, the culture
medium was removed, and the hydrogels were washed with D-PBS and fixed
with 4% paraformaldehyde. After washing with water, the hydrogel model
was permeabilized with 0.5% Triton X-100. After being washed with
D-PBS, the specimens were stained with 100 nM rhodamine-phalloidin
(Fujifilm Wako Pure Chemical Industries, Ltd., Osaka, Japan) for 40
min in the dark. After washing 3 times with D-PBS, the hydrogels were
stained with 4′,6-diamidino-2-phenylindole, dihydrochloride
(Dojindo Laboratories, Kumamoto, Japan) diluted 1:500 in D-PBS for
5 min in the dark. After washing with D-PBS, the hydrogels were observed
by using a confocal laser scanning microscope (CLSM) (FV3000, EVIDENT,
Tokyo, Japan).