EpiCult™-B Human Medium Kit

For culture and evaluation of human mammary epithelial cells in CFU assays

EpiCult™-B Human Medium Kit

For culture and evaluation of human mammary epithelial cells in CFU assays

EpiCult™-B Human Medium Kit
1 Kit
230 USD
Catalog # 05601

For culture and evaluation of human mammary epithelial cells in CFU assays

What's Included

  • EpiCult™-B Basal Medium (Human), 100 mL
  • EpiCult™-B Proliferation Supplement (Human), 1 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

EpiCult™-B Human Medium Kit is a serum-free culture medium optimized for the culture of human mammary luminal and myoepithelial cells. It is ideal for the growth and evaluation of bipotent, luminal-restricted and myoepithelial-restricted mammary epithelial progenitor cells in the mammary colony-forming unit (CFU) assay when used in conjunction with an irradiated feeder layer such as NIH 3T3.

This kit contains EpiCult™-B Basal Medium (Human) and EpiCult™-B Proliferation Supplement (Human). Addition of Hydrocortisone Stock Solution (Catalog #07925) is required.
Subtype
Specialized Media
Cell Type
Mammary Cells
Species
Human
Application
Cell Culture, Colony Assay
Brand
EpiCult
Area of Interest
Epithelial Cell Biology
Formulation
Serum-Free

Data Figures

Protocol for isolation and identification of human and mouse mammary epithelial progenitor cells

Figure 1. Protocol for isolation and identification of human and mouse mammary epithelial progenitor cells

Phase contrast photographs of (A) a pure human myoepithelial cell colony, (B) a pure human luminal cell colony, and (C) a mixed human colony. (D) is a mouse colony. Unlike human mammary CFC colonies, subtypes of mouse mammary epithelial cell colonies are not easily identifiable. All colonies were cultured in either EpiCult®-B (Human: Catalog #05601) or EpiCult®-B (Mouse:Catalog #5610) in the presence of an irradiated NIH 3T3 feeder layer. Colonies were visualized by staining with Wright"s Giemsa. (E) is a picture of mammospheres obtained from primary human mammary epithelial cells and (F) is an image of tumorspheres obtained from MCF7 human breast cancer cell line.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
05601
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
05601
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Publications (9)

Molecular decoy to the Y-box binding protein-1 suppresses the growth of breast and prostate cancer cells whilst sparing normal cell viability. Law JH et al. PloS one 2010 JAN

Abstract

The Y-box binding protein-1 (YB-1) is an oncogenic transcription/translation factor that is activated by phosphorylation at S102 whereby it induces the expression of growth promoting genes such as EGFR and HER-2. We recently illustrated by an in vitro kinase assay that a novel peptide to YB-1 was highly phosphorylated by the serine/threonine p90 S6 kinases RSK-1 and RSK-2, and to a lesser degree PKCα and AKT. Herein, we sought to develop this decoy cell permeable peptide (CPP) as a cancer therapeutic. This 9-mer was designed as an interference peptide that would prevent endogenous YB-1(S102) phosphorylation based on molecular docking. In cancer cells, the CPP blocked P-YB-1(S102) and down-regulated both HER-2 and EGFR transcript level and protein expression. Further, the CPP prevented YB-1 from binding to the EGFR promoter in a gel shift assay. Notably, the growth of breast (SUM149, MDA-MB-453, AU565) and prostate (PC3, LNCap) cancer cells was inhibited by ∼90% with the CPP. Further, treatment with this peptide enhanced sensitivity and overcame resistance to trastuzumab in cells expressing amplified HER-2. By contrast, the CPP had no inhibitory effect on the growth of normal immortalized breast epithelial (184htert) cells, primary breast epithelial cells, nor did it inhibit differentiation of hematopoietic progenitors. These data collectively suggest that the CPP is a novel approach to suppressing the growth of cancer cells while sparing normal cells and thereby establishes a proof-of-concept that blocking YB-1 activation is a new course of cancer therapeutics.
Association of reactive oxygen species levels and radioresistance in cancer stem cells. Diehn M et al. Nature 2009 APR

Abstract

The metabolism of oxygen, although central to life, produces reactive oxygen species (ROS) that have been implicated in processes as diverse as cancer, cardiovascular disease and ageing. It has recently been shown that central nervous system stem cells and haematopoietic stem cells and early progenitors contain lower levels of ROS than their more mature progeny, and that these differences are critical for maintaining stem cell function. We proposed that epithelial tissue stem cells and their cancer stem cell (CSC) counterparts may also share this property. Here we show that normal mammary epithelial stem cells contain lower concentrations of ROS than their more mature progeny cells. Notably, subsets of CSCs in some human and murine breast tumours contain lower ROS levels than corresponding non-tumorigenic cells (NTCs). Consistent with ROS being critical mediators of ionizing-radiation-induced cell killing, CSCs in these tumours develop less DNA damage and are preferentially spared after irradiation compared to NTCs. Lower ROS levels in CSCs are associated with increased expression of free radical scavenging systems. Pharmacological depletion of ROS scavengers in CSCs markedly decreases their clonogenicity and results in radiosensitization. These results indicate that, similar to normal tissue stem cells, subsets of CSCs in some tumours contain lower ROS levels and enhanced ROS defences compared to their non-tumorigenic progeny, which may contribute to tumour radioresistance.
Transcriptome analysis of the normal human mammary cell commitment and differentiation process. Raouf A et al. Cell stem cell 2008 JUL

Abstract

Mature mammary epithelial cells are generated from undifferentiated precursors through a hierarchical process, but the molecular mechanisms involved, particularly in the human mammary gland, are poorly understood. To address this issue, we isolated highly purified subpopulations of primitive bipotent and committed luminal progenitor cells as well as mature luminal and myoepithelial cells from normal human mammary tissue and compared their transcriptomes obtained using three different methods. Elements unique to each subset of mammary cells were identified, and changes that accompany their differentiation in vivo were shown to be recapitulated in vitro. These include a stage-specific change in NOTCH pathway gene expression during the commitment of bipotent progenitors to the luminal lineage. Functional studies further showed NOTCH3 signaling to be critical for this differentiation event to occur in vitro. Taken together, these findings provide an initial foundation for future delineation of mechanisms that perturb primitive human mammary cell growth and differentiation.

Contact STEMCELL Technologies

Our Customer Service, Sales, and Product and Scientific Support departments in North America are available between 6 am and 5 pm Pacific Time (9 am and 8 pm Eastern Time). One of our representatives will be happy to help you by telephone or email. Please complete the form to contact us by email. A representative will get back to you shortly.
  •  

StemCell Technologies Inc. and affiliates ("STEMCELL Technologies") does not share your email address with third parties. StemCell Technologies Inc. will use your email address to confirm your identity and send you newsletters, transaction-related emails, promotional and customer service emails in accordance with our privacy policy. You can change your email preferences at any time.