EpiCult®-B Mouse Medium Kit

Recommended for:	• Culture and evaluation of mouse mammary epithelial progenitor cells in mammary colony-forming cell assays using cells derived from primary mouse mammary gland tissue • Dissociation of mammary tissue when supplemented with collagenase and hyaluronidase
Components:	• EpiCult®-B Basal Medium (Mouse), 450mL (Catalog #05611) • EpiCult®-B Proliferation Supplement (Mouse), 50mL (Catalog #05612)
Accessory Products:	• EGF, Human, Recombinant (Catalog #02633) • bFGF, Human, Recombinant (Catalog #02634) • Heparin (Catalog #07980) • Ammonium Chloride Solution (Catalog #07800) • Collagen Solution (Bovine) (Catalog #04902) • Collagenase/Hyaluronidase Solution (Catalog #07912) • Dispase (5 mg/mL) (Catalog #07913) • DNase I (Catalog #07900) • Fetal Bovine Serum (Catalog #06100 or quality cell culture-tested equivalent) • L-glutamine (Catalgo #07100) • Hanks' Balanced Salt Solution Modified (Catalog #37150) • Tissue Dissociation Flask (Catalog #27300) • Trypsin-EDTA (0.25%) (Catalog #07901) • 40 µm Cell Strainers (Catalog #27305)
Intended Use Statement:	For Research Use Only. Not for Therapeutic or Diagnostic Use.
Product Type:	Specialized cell culture media
Application:	Colony assays & quantification
Area of Interest:	Cancer, Mammary cell research, Stem cell biology
Cell Type:	Mammary epithelial cells
Medium Type:	Liquid, Serum-free
Popular Product Line:	EpiCult
Species:	Mouse

Procedures and instruction manuals:

•  PRODUCT INFORMATION SHEET - 05610, Lot# All:29568-PIS_1_1_1.pdf

Educational resources:

•  VIDEO: MammoCult® Medium for Culture of Mammospheres and Tumorspheres
•  VIDEO: ALDEFLUOR®: Detect Normal and Cancer Precursor Cells
•  WEBINAR: ALDEFLUOR® and Cancer Stem Cells
•  BROCHURE: Products for Mouse Mammary Cell Research
•  BROCHURE: Mammary Epithelial Cells: Standardized Media and Reagents
•  MINI REVIEW: Mini-review: Mammary stem cells (Catalog #29017)
•  TECH BULLETIN: Isolation and characterization of mouse mammary stem cells (Catalog #29077)
•  BROCHURE: Standardized Solutions for Cancer Stem Cell Research
•  VIDEO: Introducing EpiCult®-C Medium

MSDS:

•  05611-MSDS.pdf
•  05612-MSDS.pdf

This product has been used in:

1. Alicia Pinderhughes et al. MMTV-Wnt1 and -DeltaN89beta-catenin induce canonical signaling in distinct progenitors and differentially activate Hedgehog signaling within mammary tumors.PLoS ONE 4 (2) e4537 (2009)
2. Daisuke Yamaji et al. Development of mammary luminal progenitor cells is controlled by the transcription factor STAT5AGenes & Development 23 (20) (October 15, 2009)
3. Yohei Shimono et al. Downregulation of miRNA-200c links breast cancer stem cells with normal stem cells.Cell 138 (3) 592-603 (August 7, 2009)
4. Norazizah Shafee et al. Cancer stem cells contribute to cisplatin resistance in Brca1/p53-mediated mouse mammary tumors.Cancer Res 68 (9) 3243-3250 (May 1, 2008)
5. J Stingl et al. Characterization of bipotent mammary epithelial progenitor cells in normal adult human breast tissue.Breast Cancer Res Treat 67 (2) 93-109 (May 2001)

Background References:

1. Mark Shackleton et al. Generation of a functional mammary gland from a single stem cell.Nature 439 (7072) 84-88 (January 5, 2006)
2. E C Kordon et al. An entire functional mammary gland may comprise the progeny from a single cell.Development 125 (10) 1921-1930 (May 1998)
3. G H Smith et al. Experimental mammary epithelial morphogenesis in an in vivo model: evidence for distinct cellular progenitors of the ductal and lobular phenotype.Breast Cancer Res Treat 39 (1) 21-31 (1996)

Product Name	Description	Catalog #
EpiCult®-B Medium Kit (Human)	Kit to Culture and Evaluate Human Mammary Epithelial Cells	05601
EpiCult®-C Human Medium Kit	Medium for Human Mammary Epithelial Cell Culture	05630
MammoCult® Human Medium Kit	Medium for the Culture of Human Mammospheres and Tumorspheres	05620
Collagenase/Hyaluronidase	10X Collagenase/Hyaluronidase in DMEM	07912
Dispase (5 mg/mL)	5 mg/mL Dispase in Hank's Balanced Salt Solution Modified	07913
DNase I Solution (1 mg/mL)	Deoxyribonuclease I Solution (1 mg/mL)	07900
Trypsin-EDTA (0.25%)	Trypsin-EDTA (0.25%)	07901
EGF, Human, Recombinant	Epidermal Growth Factor	02633
40 µm Cell Strainer	40 µm Nylon Cell Strainer	27305
Fetal Bovine Serum for Human Myeloid Colony-Forming Cells	For Preparation of Methylcellulose-Based Medium for Human Myeloid Colony-forming Cells	06100
EasySep™ Mouse Mammary Stem Cell Enrichment Kit	Immunomagnetic Negative Selection of Mouse Mammary Stem Cells (MaSC)	19757
EasySep™ Mouse Epithelial Cell Enrichment Kit	Immunomagnetic Negative Selection of Untouched Mouse Epithelial Cells	19758
FGF-8b, Mouse, Recombinant	Fibroblast growth factor 8b (FGF-8b)	02755
EGF (Carrier-Free), Human, Recombinant	Epidermal Growth Factor (Carrier Free)	02653
0.2% Heparin Sodium Salt in PBS	0.2% Heparin Sodium Salt in Phosphate Buffered Saline	07980
Mammary Stem Cell Training Course	Training Course for Mammary Stem Cells	00212

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.