Journal: Stem Cell Research & Therapy

Article Title: A robust cell culture system for large scale feeder cell-free expansion of human breast epithelial progenitors

doi: 10.1186/s13287-018-0994-y

Figure Lengend Snippet: EpiPro Plus growth media enables large-scale expansion of breast epithelial cells in 2D cultures. a Experimental outline for ex vivo propagation of primary human breast epithelial cells (HBECs) using EpiPro Plus media (Lin - , Lineage-depleted; BC, breast cells). b Top panels: representative photomicrographs of HBECs grown in 2D adherent cultures without feeder fibroblasts using EpiPro Plus media over five passages. Middle panels: representative FACS plots depicting CD49f and EpCAM expression in ex vivo expanded HBECs. Lower panels: representative photomicrographs of ex vivo expanded HBECs grown in Matrigels as organoids. Total cell numbers ( c ) and CFC yields ( d ) per passage for different ex vivo expanded cells (generated from eight different reduction mammoplasty samples) are plotted as individual line graphs. e Representative colony pictures and progenitor cell frequency (luminal and mixed colony forming cell numbers) are plotted for multiple passages. Results are represented as mean ± SEM from at least three biological replicates. Bars represent 400 μm ( b ) and 1000 μm ( e ). Representative colony pictures and progenitor cell frequencies were plotted for multiple passages. A, B gated cell populations, CFC colony forming cell, EpCAM epithelial cell adhesion molecule, Lin – lineage depleted, P passage

Article Snippet: Lineage-depleted bulk HBECs were obtained from breast reduction samples and propagated in 2D adherent cultures using the EpiPro growth medium (DMEM-F12 (with 2.5 mM l -glutamine and 15 mM HEPES) base medium supplemented with 2% (v/v) fetal bovine serum, 10 ng/ml human epidermal growth factor, 1 μg/ml insulin, 0.5 μg/ml hydrocortisone, 1% (w/v) bovine serum albumin, 10 ng/ml cholera toxin, 10 μM Y-27632 (Cat#72304; Stem Cell Tech.) and 10 μM SB-431542 (Cat#72234; StemCell Tech.)).

Techniques: Ex Vivo, Expressing, Generated

Journal: Stem Cell Research & Therapy

Article Title: A robust cell culture system for large scale feeder cell-free expansion of human breast epithelial progenitors

doi: 10.1186/s13287-018-0994-y

Figure Lengend Snippet: EpiPro Plus medium enables long-term and large-scale expansion of purified breast epithelial progenitors. a Top panels: representative photomicrographs of bipotent progenitor-enriched subset of HBECs grown in 2D adherent cultures using EpiPro Plus medium over five passages. Middle panels: representative FACS plots depicting CD49f and EpCAM expression in ex vivo expanded bipotent progenitor (CD49 bright EpCAM low )-enriched fraction. Lower panels: representative photomicrographs of ex vivo expanded bipotent progenitors grown as organoids. b Top panels: representative photomicrographs of luminal progenitors (CD49 low EpCAM bright ) grown in 2D adherent cultures using EpiPro Plus media over five passages. Middle panels: representative FACS plots depicting CD49f and EpCAM expression in ex vivo expanded luminal progenitor (CD49 low EpCAM bright ). Lower panels: representative photomicrographs of 3D organoids grown from ex vivo expanded luminal progenitors. Total cell number and CFC yield per passage calculated and plotted for ( c ) bipotent and ( d ) luminal progenitors for each individual sample. Bars represent 400 μm. Bipotent and luminal progenitors were obtained from three separate mammoplasty samples. A, B gated cell populations, CFC colony forming cell, P passage

Article Snippet: Lineage-depleted bulk HBECs were obtained from breast reduction samples and propagated in 2D adherent cultures using the EpiPro growth medium (DMEM-F12 (with 2.5 mM l -glutamine and 15 mM HEPES) base medium supplemented with 2% (v/v) fetal bovine serum, 10 ng/ml human epidermal growth factor, 1 μg/ml insulin, 0.5 μg/ml hydrocortisone, 1% (w/v) bovine serum albumin, 10 ng/ml cholera toxin, 10 μM Y-27632 (Cat#72304; Stem Cell Tech.) and 10 μM SB-431542 (Cat#72234; StemCell Tech.)).

Techniques: Purification, Expressing, Ex Vivo