Self-Renewal and Multilineage Differentiation In Vitro from Murine Prostate Stem Cells

Abstract

Murine prostate stem cells express integrin ␣6, which modulates survival, proliferation, and differentiation signaling through its interaction with the extracellular protein laminin. When plated in vitro in laminin containing Matrigel medium, 1 of 500 -1,000 murine prostate cells can grow and form clonogenic spheroid structures that we term prostate spheres. Prostate spheres can be serially passaged individually or in bulk to generate daughter spheres with similar composition, demonstrating that sphere-forming cells are capable of self-renewal. Spheres spontaneously undergo lineage specification for basal and transit-amplifying cell types. P63-expressing cells localized to the outer layers of prostate spheres possess higher self-renewal capacity, whereas cells toward the center display a more differentiated transitamplifying phenotype, as demonstrated by the expression of the prostate stem cell antigen. When dihydrotestosterone is added to the medium, the androgen receptor is stabilized, is imported to the nucleus, and drives differentiation to a luminal cell-like phenotype. A fraction of sphere cells returned to an in vivo environment can undergo differentiation and morphogenesis to form prostate tubular structures with defined basal and luminal layers accompanied by prostatic secretions. This study demonstrates self-renewal and multilineage differentiation from single adult prostate stem/ progenitor cells in a specific in vitro microenvironment.

Prostatic inflammation enhances basal-to-luminal differentiation and accelerates initiation of prostate cancer with a basal cell origin

“…The first mechanism is less likely because isolated basal cells are not able to differentiate readily into luminal or neuroendocrine cells when cultured in vitro. Hormones or defined growth factors are required for their differentiation (54)(55)(56). Therefore, future studies will focus on the identification of the basal cell-autonomous or niche-derived signaling that actively drives basal-to-luminal differentiation.…”

JNK and PTEN cooperatively control the development of invasive adenocarcinoma of the prostate

“…To test whether these cells contribute to the tumor phenotype, we isolated lineage-negative (Lin − ) Sca1 + cells from ΔJnk ΔPten and ΔPten primary tumors and cultured these cells in vitro. Previous studies using WT mice have demonstrated that this procedure leads to the formation of spheres (prostaspheres) with a surface location of prostate stem cells and a luminal location of more differentiated cells (31). Luminal prostate stem cells may also be present in these cultures (32,33).…”

Stem‐Like Epithelial Cells Are Concentrated in the Distal End of the Fallopian Tube: A Site for Injury and Serous Cancer Initiation

“…The conditions to culture and passage spheres were adapted based on previously described protocols [18]. In short, FACSisolated epithelial single-cell preparations were suspended in equal volumes of PrEGM media (Lonza CC-3166, Allendale, NY, http://www.lonza.com) and Matrigel (BD Biosciences).…”