Abstract
The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution. Mammary gland tumors represent the most common cancer diagnosed in women worldwide. Studying the regulatory mechanisms of mammary gland development is essential for understanding how dysregulation can lead to breast cancer initiation and progression. Three-dimensional (3D) mammary organoids offer many exciting possibilities for the study of tissue development and breast cancer. In the present protocol derived from Sumbal et al., we describe a straightforward 3D organoid system for the study of lactation and involution ex vivo. We use primary and passaged mouse mammary organoids stimulated with fibroblast growth factor 2 (FGF2) and prolactin to model the three cycles of mouse mammary gland lactation and involution processes. This 3D organoid model represents a valuable tool to study late postnatal mammary gland development and breast cancer, in particular postpartum-associated breast cancer. Graphic abstract: Mammary gland organoid isolation and culture procedures.
Highlights
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[Abstract] The mammary gland is a highly dynamic tissue that changes throughout reproductive life, including growth during puberty and repetitive cycles of pregnancy and involution
In the present protocol derived from Sumbal et al, we describe a straightforward 3D organoid system for the study of lactation and involution ex vivo
Summary
Dissection of a virgin mouse to harvest mammary glands (see Video 1). This video was made at Pasteur Institute. A. Euthanize the donor mouse using an ethically approved method (e.g., cervical dislocation) and immediately proceed to mammary gland collection. Cervical dislocation is a common method for animal euthanasia and provides a fast and painless death With this method, cell/tissue survival in culture is not altered if collected immediately. From this median line, use forceps to grasp the skin and cut toward each of the four paws (see Figure 1A, incise to join the middle line to points C, D, E or F, respectively). Using curved forceps, grasp the mammary glands and gently separate them from the skin and other tissues with scissors. Dispose of the animal corpse and continue with mechanical and enzymatic dissociation of the mammary glands
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