Mammospheres, or mammary epithelial stem cell aggregates, derived from primary breast tumors or cell lines are thought to develop from rare cancer stem cell (CSC) subpopulations within the tumor. The progression and recurrence of breast cancer, the leading cause of cancer death in women, has been linked to small populations of cancer stem cells (CSCs).1 The human mammary gland can undergo multiple cycles of changes in prooliferation and function controlled by stem cells. Signaling pathways involved in mammary stem cell regulation are often deregulated in breast cancer.1 These cancer stem cell populations express stem cell related markers Nestin and Sox2 while lacking GFAP (2). The 3D culture of breast-derived cancer cells is considered to be a more biologically relevant cell culture model maintaining the original malignant tumor phenotype.
Breast cancer cells cultured in fetal bovine serum (FBS) undergo unwanted phenotype changes and cell differentiation, while culturing 3D mammospheres in serum-free conditions maintains the original properties of the primary tumor.4 However, conventional serum-free media for culturing mammospheres struggle to efficiently maintain the self-renewing subpopulations of cancer stem cells (CSCs). Consequently, the stem cell population is gradually lost overtime along with the cell proliferation rates. The PromoCell 3D Tumorsphere Media XF (C-28070, C-28075) is designed to serially passage breast cancer derived cells as undifferentiated 3D mammospheres with high cell proliferation rates. It’s defined and serum-free formualtion allows consistent and standardized expansion of mammospheres while maintaining the cancer stem cell characteristics such as self-renewal and chemoresistance.
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Passage the spheroid before they form a dark necrotic center (typically 4-10 days). Collect
Figure 1. Cell proliferation rates of MCF-7 and MDA-MB-231 breast cancer cells during 3D mammosphere culture. 4X104 MCF-7 (86012803) or MDA-MB-231 (92020424) cells/well were plated in triplicate using 6-well ULA suspension plates. Serial passage by enzymatic dissociation was performed according to protocol. Mammosphere formation and cell proliferation were maintained during 3D culture over 11 passages.
Figure 2. Mammosphere culture of MCF-7 and MDA-MB-231 breast cancer cells in PromoCell 3D Tumorsphere Media XF. Mammospheres were serially passaged by enzymatic dissociation was performed according to protocol. Mammosphere formation and cell proliferation were maintained during 3D culture over 11 passages.
Figure 3. Population doublings and tumor formation efficiencies of MCF-7 breast cancer cells during 3D mammosphere culture.A) 4X104 MCF-7 cells/well were plated in triplicate using 6-well ULA suspension plates. Serial passage by enzymatic dissociation was performed every 9 days. Consistent mammosphere formation and cell proliferation were maintained during 3D culture over 11 passages. B) Serial passage of MCF-7 cells over 9 passages results in significant increase in tumor formation efficiency from 2% to 28%.
Figure 4. Cell morghology of MDA-MB-231 breast cancer cells during 3D mammosphere culture.Mammospheres form non-adherent suspension aggregates in PromoCells’s Tumorsphere Media XF compared to unwanted adherent growth pattern in competitors tumorpshere media.