세포주 정보

15, 16

18

22

X

8

8

9, 10

7, 11

11

11

DMEM/F-12, 52.5%; FBS, 40%; DMSO, 7.5%

50% Advanced DMEM/F12, 40% WRN CM, 5% FBS, 1 x GlutaMAX, 1 x B27, 10mM Nicotinamide, 20ng/mL human EGF, 10ng/mL human FGF-10, 10ng/mL IGF-1, 10ng/mL Heregulinβ-1, 10nM β-estradiol,1.25mM N-acetyl cysteine, 5uM A83-01,10uM Y-27632, 50 µg/mL primocin

50% Advanced DMEM/F12, 40% WRN CM, 5% FBS, 1 x GlutaMAX, 1 x B27, 10mM Nicotinamide, 20ng/mL human EGF, 10ng/mL human FGF-10, 10ng/mL IGF-1, 10ng/mL Heregulinβ-1, 10nM β-estradiol,1.25mM N-acetyl cysteine, 5uM A83-01,10uM Y-27632, 50 µg/mL primocin

The organoid-containing dome (Corning, Matrigel; Trevigen, Cultrex BME; Merck, ECM; Thermofisher, Geltrex) was mechanically pipetted using animal origin-free, recombinant enzyme (Thermofisher사 TrypLE Express solution) and organoids were collected in a 15 mL conical tube. The organoid-containing dome was mechanically dissociated with intense pipetting, and the tube containing the organoids and organoid-containing dome mixture was incubated at 37°C for approximately 5 - 10 min. The organoids were centrifuged at 1,000 rpm for 3 min, and the supernatant was aspirated. Once organoid-containing dome was removed, the cell pellet was resuspended with fresh organoid-containing dome, seeded in a T-25 flask and the flask was incubated at 37℃ for 10 min. Once organoid-containing dome was solidified, 5 mL of the culture medium was added to the flask to overlay the organoid-containing dome and cells were incubated in a 37℃ and 5% CO2 culture incubator.

HPV16, E1 gene, deleted, E2 gene, deleted by PCR analysis; HPV 16 was presumably integrated into chromosome; HPV16 was not detectable by Southern blot analysis; E6/E7 gene of HPV16 was expressed by Northern analysis., No antibiotics were added to culture media (No penicillin and streptomycin)

Medium is aspirated. Care was taken not to destroy the BME dome. 5 mL of medium was added, and cells were incubated in a 37℃ and 5% CO2 culture incubator.