Using small intestinal epithelial cells, the organoid culture was performed as described previously 33 (link). Briefly, the organoids were cultured in matrigel with advanced DMEM/F12 medium (Invitrogen) supplemented with 50 ng/mL EGF (Invitrogen), R-Spondin1 conditioned medium (R&D Systems), and 100 ng/mL Noggin (Peprotech). The culture was passaged once, and the crypt length was measured under a dissecting microscope. Organoid cell proliferation was detected using the Click-iT EdU Imaging System (Invitrogen).
R spondin1 conditioned medium
Manufactured by R&D Systems
Sourced in United States
R-Spondin1 conditioned medium is a cell culture supplement produced by R&D Systems. It contains secreted R-Spondin1 protein, which is a key regulator of the Wnt signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Click it edu imaging system, by Thermo Fisher Scientific (2 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (2 mentions)
Advanced dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Noggin, by Thermo Fisher Scientific (1 mentions)
Y 27632, by AbMole (1 mentions)
B27 supplement, by Thermo Fisher Scientific (1 mentions)
Gastrin 1, by Bio-Techne (1 mentions)
Wnt3a conditioned medium, by R&D Systems (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
A83 01, by Bio-Techne (1 mentions)
Glutamax 1, by Thermo Fisher Scientific (1 mentions)
Fgf10, by Thermo Fisher Scientific (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Nicotinamide, by Merck Group (1 mentions)
3 protocols using r spondin1 conditioned medium
1
Intestinal Epithelial Organoid Culture
2
Organoid Culture Medium Composition
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The organoid culture medium consisted of Advanced DMEM/F12, Primocin (100 μg/mL, Invitrogen)，HEPES (10 mM, Gibco), GlutaMAX‐I (1X, Gibco), A83‐01 (500 nM, Tocris), Y‐27632 (10 μM, AbMole), N‐acetylcysteine (1.56 mM, Sigma), nicotinamide (10 mM, Sigma), FGF10 (10 ng/mL, PeproTech), B27supplement (1X, Gibco), Epidermal Growth Factor (50 ng/mL, PeproTech), Gastrin I (0.01 μM, Tocris Bioscience), Wnt3A conditioned medium (50 ng/mL, R&D Systems)，R‐spondin‐1 conditioned medium (250 ng/mL, R&D Systems), and Noggin conditioned medium (100 ng/mL, R&D Systems).
3
Organoid Cultivation from Intestinal Cells
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The cultivation of organoids from small intestinal epithelial cells was conducted following previously established protocols [25 (link)]. In summary, the resulting organoids were maintained within Matrigel, employing advanced DMEM/F12 medium (HY-K3002; Sigma) that was supplemented with 50 ng/mL epidermal growth factor (EGF) (HY-P70590AF; Sigma), R-Spondin1 conditioned medium (R&D Systems, Minneapolis, MN, USA), and 100 ng/mL Noggin (250-38-1MG; PeproTech, Cranbury, NJ, USA). The cultures underwent a single passage, with crypt length measurements performed under a dissecting microscope. The proliferation of organoid cells was assessed using the Click-iT EdU Imaging System (Invitrogen).
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