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. 2022 Mar 16;13:826107. doi: 10.3389/fphys.2022.826107

Figure 1.

Figure 1

Organoid culture protocol and timeline. (A-C) Dissection, digestion, and centrifugation steps to isolate mammary epithelial fragments for organoid culture. (D) Organoid expansion step in BME to generate mammary spheres. (E) Collagen pre-assembly, pH adjusting step, and generation of BME:collagen I mixture (7:3) to plate mammary spheroids in 7B3C. (F) Alternation treatment with FGF2 and EGF to generate branched mammary organoids. (G) Timeline of mammary gland harvesting procedure and processing steps to obtain branched organoids in 7B3C gels using alternating treatment conditions.

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