MammoCult™ Human Medium Kit

For culture of human mammospheres and tumorspheres
MammoCult™ Human Medium Kit

For culture of human mammospheres and tumorspheres

1 Kit
Catalog # 05620
313 USD
Required Products
  1. EpiCult™-B Medium Kit (Human)
    EpiCult™-B Human Medium Kit

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

  2. Heparin Solution
    Heparin Solution

    Cell culture supplement

  3. 37μm Reversible Strainers|27215,27250
    Reversible Strainers

    Reversible strainer for filtration of single cells and isolation of cellular aggregates, including embryoid bodies and spheroids

  4. 6-Well Ultra-Low Adherent Plate|38071
    Ultra-Low Adherent Plate for Suspension Culture

    Sterile, flat-bottom plate, with ultra-low adherent surface for suspension cultures, with lid; 6-well format

Overview

MammoCult™ Medium (Human) is a serum-free culture medium optimized for the culture of mammospheres from normal human primary breast tissues and tumorspheres from human breast cancer cell lines. For preparation of complete MammoCult™ Medium, Hydrocortisone Stock Solution (Catalog #07925) and Heparin Solution (Catalog #07980) are also required.
Components
  • MammoCult™ Basal Medium (Human), 450 mL
  • MammoCult™ Proliferation Supplement (Human), 50 mL
Subtype
Specialized Media
Cell Type
Cancer Cells and Cell Lines, Mammary Cells
Species
Human
Application
Cell Culture, Maintenance, Spheroid Culture
Brand
MammoCult
Area of Interest
Cancer, Epithelial Cell Biology
Formulation
Serum-Free

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
MammoCult™ Human Medium Kit
Catalog #
05620
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
MammoCult™ Human Medium Kit
Catalog #
05620
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
MammoCult™ Human Medium Kit
Catalog #
05620
Lot #
All
Language
English

Educational Materials (12)

Brochure
Mammary Epithelial Cells: Standardized Media and Reagents
Technical Bulletin
Tumorsphere Culture of Human Breast Cancer Cell Lines
Wallchart
SnapShot: Breast Cancer
Video
Culture Mammospheres and Tumorspheres with the MammoCult™ Medium
0:51
Culture Mammospheres and Tumorspheres with the MammoCult™ Medium
Video
How to Count 3D Sphere Cultures (Mammospheres, Neurospheres, Cancer Spheroids, and PSC Aggregates)
2:05
How to Count 3D Sphere Cultures (Mammospheres, Neurospheres, Cancer Spheroids, and PSC Aggregates)
Video
How to Detect and Isolate Normal and Cancer Stem Cells Based on ALDH Activity with ALDEFLUOR™
1:22
How to Detect and Isolate Normal and Cancer Stem Cells Based on ALDH Activity with ALDEFLUOR™
Video
Drug Screening and Phenotypic Analysis in a Microwell-based 3D Cell Culture System
6:31
Drug Screening and Phenotypic Analysis in a Microwell-based 3D Cell Culture System
Video
Tools for Breast Cancer Research
1:23
Tools for Breast Cancer Research
Video
How to Generate Uniform, Size-Controlled 3D Spheroid Cultures with AggreWell™ Microwell Plates
0:52
How to Generate Uniform, Size-Controlled 3D Spheroid Cultures with AggreWell™ Microwell Plates
Webinar
Potential Applications of ALDH Bright Cells In Regenerative Medicine
53:18
Potential Applications of ALDH Bright Cells In Regenerative Medicine
Scientific Poster
Mammosphere Culture Supports Short But Not Long-Term Propagation of Human Mammary Epithelial Progenitors
Scientific Poster
Drug Screening and Phenotypic Analysis in a Microwell-based 3D Cell Culture System

Product 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.

Data and Publications

Data

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.

Publications (85)

British journal of cancer 2016 MAY Regulation of the T-box transcription factor Tbx3 by the tumour suppressor microRNA-206 in breast cancer. Amir S et al.

Abstract

BACKGROUND The Tbx3 transcription factor is over-expressed in breast cancer, where it has been implicated in proliferation, migration and regulation of the cancer stem cell population. The mechanisms that regulate Tbx3 expression in cancer have not been fully explored. In this study, we demonstrate that Tbx3 is repressed by the tumour suppressor miR-206 in breast cancer cells. METHODS Bioinformatics prediction programmes and luciferase reporter assays were used to demonstrate that miR-206 negatively regulates Tbx3. We examined the impact of miR-206 on Tbx3 expression in breast cancer cells using miR-206 mimic and inhibitor. Gene/protein expression was examined by quantitative reverse-transcription-PCR and immunoblotting. The effects of miR-206 and Tbx3 on apoptosis, proliferation, invasion and cancer stem cell population was investigated by cell-death detection, colony formation, 3D-Matrigel and tumorsphere assays. RESULTS In this study, we examined the regulation of Tbx3 by miR-206. We demonstrate that Tbx3 is directly repressed by miR-206, and that this repression of Tbx3 is necessary for miR-206 to inhibit breast tumour cell proliferation and invasion, and decrease the cancer stem cell population. Moreover, Tbx3 and miR-206 expression are inversely correlated in human breast cancer. Kaplan-Meier analysis indicates that patients exhibiting a combination of high Tbx3 and low miR-206 expression have a lower probability of survival when compared with patients with low Tbx3 and high miR-206 expression. These studies uncover a novel mechanism of Tbx3 regulation and identify a new target of the tumour suppressor miR-206. CONCLUSIONS The present study identified Tbx3 as a novel target of tumour suppressor miR-206 and characterised the miR-206/Tbx3 signalling pathway, which is involved in proliferation, invasion and maintenance of the cancer stem cell population in breast cancer cells. Our results suggest that restoration of miR-206 in Tbx3-positive breast cancer could be exploited for therapeutic benefit.
Nanomedicine (London, England) 2016 MAY Evaluation of expansile nanoparticle tumor localization and efficacy in a cancer stem cell-derived model of pancreatic peritoneal carcinomatosis. Herrera VL et al.

Abstract

AIM To evaluate the tumor localization and efficacy pH-responsive expansile nanoparticles (eNPs) as a drug delivery system for pancreatic peritoneal carcinomatosis (PPC) modeled in nude rats. METHODS & MATERIALS A Panc-1-cancer stem cell xeno1graft model of PPC was validated in vitro and in vivo. Tumor localization was tracked via in situ imaging of fluorescent eNPs. Survival of animals treated with paclitaxel-loaded eNPs (PTX-eNPs) was evaluated in vivo. RESULTS The Panc-1-cancer stem cell xenograft model recapitulates significant features of PPC. Rhodamine-labeled eNPs demonstrate tumor-specific, dose- and time-dependent localization to macro- and microscopic tumors following intraperitoneal injection. PTX-eNPs are as effective as free PTX in treating established PPC; but, PTX-eNPs result in fewer side effects. CONCLUSION eNPs are a promising tool for the detection and treatment of PPC.
The Biochemical journal 2016 MAY Selenophosphate Synthetase 1 is an Essential Protein with Roles in Regulation of Redox Homeostasis in Mammals. Tobe R et al.

Abstract

Selenophosphate synthetase (SPS) was initially detected in bacteria and was shown to synthesize selenophosphate, the active selenium donor. However, mammals have two SPS paralogs, which are designated SPS1 and SPS2. Although it is known that SPS2 catalyzes the synthesis of selenophosphate, the function of SPS1 remains largely unclear. To examine the role of SPS1 in mammals, we generated a Sps1 knockout mouse and found that systemic SPS1 deficiency led to embryos that were clearly underdeveloped by E8.5 and virtually resorbed by E14.5. The knockout of Sps1 in the liver preserved viability, but significantly affected the expression of a large number of mRNAs involved in cancer, embryonic development, and the glutathione system. Particularly notable was the extreme deficiency of glutaredoxin 1 (GLRX1) and glutathione-S-transferase omega 1. To assess these phenotypes at the cellular level, we targeted the removal of SPS1 in F9 cells, a mouse embryonal carcinoma cell line, which affected the glutathione system proteins and accordingly led to the accumulation of hydrogen peroxide in the cell. Further, we found that several malignant characteristics of SPS1-deficient F9 cells were reversed, suggesting that SPS1 played a role in supporting and/or sustaining cancer. In addition, the overexpression of mouse or human GLRX1 led to a reversal of observed increases in reactive oxygen species (ROS) in the F9 SPS1/GLRX1-deficient cells and resulted in levels that were similar to those in F9 SPS1-sufficient cells. The results suggested that SPS1 is an essential mammalian enzyme with roles in regulating redox homeostasis and controlling cell growth.
Journal of cellular biochemistry 2016 JUN Preservation of the 3D Phenotype Upon Dispersal of Cultured Cell Spheroids into Monolayer Cultures. Koshkin V et al.

Abstract

In functional cytometric studies, cultured cells are exposed to effectors (e.g. drugs), and the heterogeneity of cell responses are studied using cytometry techniques (e.g. image cytometry). Such studies are difficult to perform on 3D cell cultures. A solution is to disperse 3D clusters and transfer the cells to the 2D state before applying effectors and using cytometry. This approach requires that the lifetime of the 3D phenotype be longer than the duration of the experiment. Here we studied the dynamics of phenotype transformation from 3D to 2D and searched for means of slowing this transformation down in dispersed spheroids of MCF7 cells. We found three functional biomarkers of the 3D phenotype in MCF7 cell spheroids that are absent in the 2D cell culture: (i) the presence of a subpopulation with an elevated drug-expelling capacity, (ii) the presence of a subpopulation with an elevated cytoprotective capacity and (iii) the accumulation of cells in the G1 phase of the cell cycle. Monitoring these biomarkers in cells transferred from the 3D state to the 2D state revealed their gradual extinction. We found that the combined application of an elevated cell density and thiol-containing medium supplements increased the lifetime of the 3D phenotype by several fold to as long as 96 h. Our results suggest that extending the lifetime of the 3D phenotype in the cells transferred from the 3D state to the 2D state can facilitate detailed functional cytometric studies, such as measurements of population heterogeneity of cytotoxicity, chemosensitivity and radiosensitivity. This article is protected by copyright. All rights reserved.
Molecular cancer research : MCR 2016 JAN Tumor Cell-Derived Periostin Regulates Cytokines That Maintain Breast Cancer Stem Cells. Lambert AW et al.

Abstract

UNLABELLED Basal-like breast cancer (BLBC) is an aggressive subtype of breast cancer which is often enriched with cancer stem cells (CSC), but the underlying molecular basis for this connection remains elusive. We hypothesized that BLBC cells are able to establish a niche permissive to the maintenance of CSCs and found that tumor cell-derived periostin (POSTN), a component of the extracellular matrix, as well as a corresponding cognate receptor, integrin $$(v)$$(3), are highly expressed in a subset of BLBC cell lines as well as in CSC-enriched populations. Furthermore, we demonstrated that an intact periostin-integrin $$3 signaling axis is required for the maintenance of breast CSCs. POSTN activates the ERK signaling pathway and regulates NF-$$B-mediated transcription of key cytokines, namely IL6 and IL8, which in turn control downstream activation of STAT3. In summary, these findings suggest that BLBC cells have an innate ability to establish a microenvironmental niche supportive of CSCs. IMPLICATIONS The findings reported here indicate that POSTN produced by CSCs acts to reinforce the stem cell state through the activation of integrin receptors and the production of key cytokines.
Phytomedicine : international journal of phytotherapy and phytopharmacology 2016 JAN Rhodiola crenulata induces an early estrogenic response and reduces proliferation and tumorsphere formation over time in MCF7 breast cancer cells. Bassa LM et al.

Abstract

BACKGROUND Rhodiola crenulata is a Tibetan mountainous plant, commonly used in Eastern alternative medicine. Many phytochemicals possess estrogenic activity, a critical regulator of proliferation in mammary epithelial cells. We have previously characterized anti-cancer properties of R. crenulata in aggressive triple negative breast cancer cells, lacking the expression of estrogen receptor. Currently, it is unknown whether R. crenulata exerts estrogenic effects and as such consumption may be a concern for women with estrogen receptor positive breast cancer that use Rhodiola sp. to relieve mild to moderate depression. PURPOSE In this study, we wished to determine whether a hydroalcoholic fraction of the R. crenulata root extract exhibits estrogenic activity in estrogen receptor positive (ER+) breast cancer cells in vitro and whether it affects normal mammary epithelial ER target gene expression in vivo. METHODS ER transcriptional activity was analyzed in MCF7 cells expressing an ERE reporter construct and confirmed via qPCR of endogenous ER target genes. We also monitored cellular proliferation over time. Additionally, to assess stem-like properties in MCF7 cells, we performed a tumorsphere formation assay under anchorage independent conditions. We examined whether R. crenulata treatment reduced $$-catenin levels via Western blotting and measured $$-catenin transcriptional activity by a reporter assay. To examine the effects of R. crenulata on normal mammary epithelial cells, we performed immunohistochemical staining of ER and PR in the mammary glands of mice fed R. crenulata for 12 weeks. RESULTS We show an initial activation of ER transcriptional activity by dual reporter assay, qPCR and proliferation of MCF7 ER+ cells in response to 24 h of R. crenulata treatment. However, upon longer treatment basal and R. crenulata induced transcriptional activity was suppressed. There was a decrease in cell doubling times and a decrease in tumorsphere formation. In association with these changes, ER$$ transcript levels were decreased and active $$-catenin levels were reduced in the cells treated for 2 weeks. Finally, we show no change in estrogen targets in normal mammary cells in vivo. CONCLUSION These data suggest that the R. crenulata extract contains components with estrogenic activity. However, R. crenulata treatment could still be protective in ER+ breast cancer cells, as longer treatment reduced the transcriptional activity of $$-catenin and ER responses leading to reduced proliferation and tumorsphere formation. Furthermore, administration of 20 mg/kg/day R. crenulata to mice did not have an observable effect on mammary epithelial ER$$ target gene expression in vivo.
View All Publications

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