Mice were sacrificed and the small intestine was collected. After a first wash in PBS buffer, the intestines were cut into pieces and dissociated with 2 mM EDTA buffer in 15 mL centrifuge tubes for 20 min at 4 °C. The samples were centrifuged at 300× g for 5 min to remove the EDTA buffer, and repeatedly pipetted with 5 mL PBS buffer. Next, crypts were collected through a 70 µm strainer (BD Biosciences, San Jose, CA, USA). Single crypts were centrifuged at 300× g for 5 min, and cultured in a 24-well tissue culture plate (Corning, New York, NY, USA) with Matrigel Matrix (Corning, New York, NY, USA) and Murine Intestinal Organoids Growth Medium (STEMCELL Technologies, Vancouver, BC, Canada). For the RANKL stimulation assay, organoids were stimulated with 100 ng/mL recombinant human RANKL (CST, Fall River, MA, USA) for one day (PBS buffer was used as control). The RANKL and mock control reagents were added daily.
Murine intestinal organoids growth medium
Manufactured by STEMCELL
Sourced in United States
Murine Intestinal Organoids Growth Medium is a specialized culture medium designed to support the growth and maintenance of murine intestinal organoids in vitro. The medium provides the necessary nutrients and growth factors to enable the proliferation and differentiation of intestinal stem cells into organoid structures that recapitulate the architecture and function of the native intestinal epithelium.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using murine intestinal organoids growth medium
1
Isolation and Culture of Intestinal Organoids
2
Intestinal Organoid RANKL Stimulation
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Mice were sacri ced to harvest the small intestines. After being washed by PBS buffer, the intestines were cut into pieces and dissociated with 2 mM EDTA buffer in 15 mL centrifuge tubes for 20 min at 4°C. The samples were centrifuged at 300 ×g for 5 min to remove the EDTA buffer, repeatedly pipetted with 5 mL PBS buffer. The crypts were then collected through a 70 µm strainer (BD, U.S.), centrifuged at 300 ×g for 5 min, and cultured in the 24-well tissue culture plate (Corning, U.S.) with Matrigel Matrix (Corning, U.S.) and Murine Intestinal Organoids Growth Medium (STEMCELL, Canada). For the RANKL stimulation assay, organoids were stimulated with 100 ng/mL recombinant human RANKL (CST, U.S.) for one day (PBS buffer was used as control). The RANKL and mock control reagents were added daily.
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