Figure 1. EpiCult™ Plus Enables the Long-Term Growth of Epithelial Stem Cells From Prostate and Mammary Tissues

(A) Mouse mammary, (B) mouse prostate, and (C) human prostate samples were established and expanded in EpiCult™ Plus. Growth curves from three independent samples are shown here.

Figure 2. Human Mammary Epithelial Cells Expand More Rapidly When Cultured in EpiCult™ Plus Compared to Conventional Epithelial Cell Expansion Media

Human mammary epithelial cells expanded in EpiCult™ Plus (solid line, solid markers) can be grown beyond 20 population doublings while cells grown in conventional epithelial cell expansion medium (dotted line, solid markers) senesced after approximately 10 population doublings. The addition of FBS to EpiCult™ Plus (solid line, open markers) allowed further expansion of the human mammary epithelial cells.

Figure 3. Cells Cultured in EpiCult™ Plus are Maintained in a Basal-Like State

EpiCult™ Plus maintains epithelial stem cells in a basal-like state as observed by the marker expression of CD49f in (A) mouse mammary, (B) mouse prostate, (C) human mammary, and (D) human prostate epithelial cells that were cultured in EpiCult™ Plus for a minimum of 8 passages. Flow cytometry analysis shows > 99% expression of CD49f across tissues.

Figure 4. Mouse Prostate and Mammary Epithelial Cells Expanded with EpiCult™ Plus Can Be Differentiated as Organoids

Mouse epithelial cells were expanded in EpiCult™ Plus and differentiated into either (A,B) prostate or (C,D) mammary organoids. (B,D) Both mouse prostate and mammary organoids express the luminal marker keratin 8 (green) and the basal marker keratin 14 (red) as shown by immunofluorescence microscopy; nuclei are counterstained with DAPI. This data demonstrates the compatibility of EpiCult™ Plus with both mouse prostate and mammary workflows that may require the robust upstream expansion of epithelial cells.

Figure 5. Human Prostate Epithelial Cells Expanded in EpiCult™ Plus Can Be Differentiated as Organoids

Freshly dissociated human prostate cells were (A) seeded and expanded in EpiCult™ Plus. After 5 days in culture, the cells were harvested and seeded into human prostate organoid medium and maintained for an additional (B) 11 days. (C) The resulting organoids contain a cell-dense core composed of luminal cells expressing keratin 8 (green) and androgen receptors (magenta), while basal cells expressing keratin 14 (red) line the outer side of the prostate organoid. This demonstrates that EpiCult™ Plus supports the robust expansion of human epithelial prostate organoids.