Gibco™Advanced DMEM/F-12

3D Cell Culture Media Human liver organoids

Experiment
3D Cell Culture Media Human liver organoids
Product
Gibco™Advanced DMEM/F-12 from Thermo Fisher Scientific
Manufacturer
Thermo Fisher Scientific

Protocol tips

Upstream tips
Defrost in advance the BME gel before use
Protocol tips
Split primary organoids after 14 days and for maintenance every 7-10 days

Publication protocol

Tissue processing, cell isolation and organoid culture. Liver cells were dissociated from the human biopsy by collagenase digestion. Cells were washed with cold Advanced DMEM/F12, filtered with a 70 μm filter and the cell pellet resuspendend in BMER (Amsbio). Isolation medium was based on AdDMEM/F12 (Invitrogen) supplemented with: 1X B27 (both from GIBCO), 1.25 mM N-Acetylcysteine (Sigma), 10 nM gastrin (Sigma), 50 ng/ml EGF (Peprotech), 10% RSPO1 conditioned media (homemade), 100 ng/ml FGF10 (Peprotech), 25 ng/ml HGF (Peprotech), 10 mM Nicotinamide (Sigma), 5 μM A83.01 (Tocris), 10 μM FSK (Tocris), 25 ng/ml Noggin (Peprotech), 30% Wnt conditioned media (homemade), and 10 uM Y27632 (Sigma Aldrich). After 3-4 days, the isolation medium was changed into a medium without Noggin, Wnt conditioned media, Y27632. Organoids could be splitted after ca. 14 days by mechanical shearing and re-plated into fresh BMER. Passage was performed in a 1:5 split ratio once every 7–10 days for at least 6 months.

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Discussion

Discussion

3 years ago

Author:

Impact of using FBS

I work with Human liver organoids and I would like to know the impact of FBS on the organoids. Since the composition of FBS is unknown, would you recommend any alternatives such as human platelet lysate?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing 3D Cell Culture Media Human liver organoids using Gibco™Advanced DMEM/F-12 from Thermo Fisher Scientific.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Gibco™Advanced DMEM/F-12 below.

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