Have questions? 1.800.667.0322  | Email Us

MethoCult™ H4100

Base Methylcellulose Medium for Human Cells

  • MethoCult™ H4100 40 mL
  • Label for MethoCult™ H4100 40 mL
MethoCult™ H4100 40 mL
Product Catalog # Size Price
MethoCult™ H4100
Base methylcellulose medium for human cells
04100 40 mL 76.00 USD
 

Related Products

Order Offline:

Call Us: +1.604.877.0713  
Toll Free: 1.800.667.0322  
Email Us:
 
Fax Us: +1.604.877.0704
Toll Free Fax: 1.800.567.2899

Quick Order

  • MethoCult™ H4100 (04100)
    40 mL
    76.00 USD

Description


MethoCult™ H4100 is a base methylcellulose medium for the culture of human cells in semisolid media. The medium is supplied as a 2.6% solution of methylcellulose in Iscove's MDM, allowing the addition of serum, growth factors and other supplements for colony-forming cell (CFC) assays of human cells. The medium can also be used as a base for cloning cell lines in semisolid media.
Application: Colony assays & quantification
Product Type: Specialized cell culture media
Recommended For:
• Assays of human clonogenic hematopoietic progenitor cells
• Testing sera, erythropoietin and cytokines
• Cloning of cell lines
Accessory Products:
• Iscove's Modified Dulbecco's Medium (IMDM) with 2% Fetal Bovine Serum (Catalog #07700)
• Blunt-end Needles (Catalog #28110/28120)
• 3 cc Syringe (Catalog #28230/28240)
• 35 mm Culture Dishes (Catalog #27100/27150)
• 60 mm Gridded Scoring Dishes (Catalog #27500)
Contains:
2.6% Methylcellulose in Iscove's MDM
Area of Interest: Cancer, Hematologic malignancies, Hematopoietic stem cell research, Stem cell biology
Cell Type: Hematopoietic stem & progenitor cells
Medium Type: Methylcellulose-based
Popular Product Line: MethoCult™
Species: Human
Intended Use Statement: FOR RESEARCH USE ONLY. NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES.
ISO Statement: STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485 MEDICAL DEVICE STANDARDS

Technical Resources


Protocols and Manuals

Current documents
Product NameLot # ?Document TypeDownload
MethoCult™ H4100 All Manual
MethoCult™ H4100 All Product Information Sheet

MSDS

Educational Resources

Videos & Webinars

  • Procedure for Setting Up the CFC Assay
  • Brochures

  • Your Ideas. Our Tools. Products for every step of your hematopoietic stem and progenitor cell research.

  • FAQS


      • Q. WHY USE SEMI-SOLID MEDIA?
        A. Semi-solid media (such as methylcellulose-based or collagen-based) allow the clonal progeny of a single progenitor cell to stay together so you can recognize distinct colonies.
      • Q. WHY USE METHYLCELLULOSE-BASED MEDIA?
        A. Methylcellulose is now widely used as a gelling agent because it permits better growth of erythroid colonies than other types of semi-solid support systems (e.g. agar) while allowing optimal granulocyte/macrophage colony formation. Committed progenitors for both erythroid and granulocyte/macrophage lineages, as well as multi-potential progenitors, can therefore be assayed simultaneously in the same culture dish.
      • Q. IS IT NECESSARY TO ADD ANTIBIOTICS TO THE MEDIA?
        A. No, we do not use antibiotics in our lab for colony assays. Aseptic technique should suffice. However, Penicillin/Streptomycin and an anti-fungal drug like amphotericin B can be added to the methylcellulose. The media should be mixed well prior to the addition of cells. Presence of antibiotics does not inhibit progenitor growth.
      • Q. IS THERE ANYTHING I CAN DO IF MY PLATES ARE CONTAMINATED?
        A. No, once contamination is visible, it is not possible to rescue the cultures by the addition of antibiotics. The bacteria/yeast inhibit colony formation by depleting nutrients or releasing toxic substances.
      • Q. WHY CAN'T I USE A PIPETTE TO DISPENSE METHYLCELLULOSE-BASED MEDIA?
        A. Because methylcellulose is a viscous solution and cannot be accurately dispensed using pipettes due to adherence of the medium to the walls of the pipette. 16-gauge (blunt-end) needles are recommended for accurate dispensing.
      • Q. CAN I 'PLUCK' THE COLONIES?
        A. Yes, colonies can be 'plucked' using a pipettor and 200 µL sterile pipette tips or glass Pasteur pipette with an elongated tip. Colonies should be placed in a volume of 25 - 50 µL and diluted into suitable culture medium.
      • Q. IS THERE A METHOCULT™ FORMULATION SUITABLE FOR HPP-CFC (HIGH PROLIFERATIVE POTENTIAL COLONY FORMING CELL)?
        A. Yes, MethoCult™ H4535 can be used as HPP-CFC does not require EPO. The culture period is usually 28 days. Feeding the media is not required as the growth factors are present in excess in the media. As these colonies can be quite large, overplating can be a problem. It is recommended to plate two different concentrations. The standard concentration, as well as a lower concentration.
      • Q. WHY ARE LOW ADHERENCE DISHES SO IMPORTANT?
        A. Because adherent cells such as fibroblasts can cause inhibition of colony growth and obscure visualization of colonies.
      • Q. CAN METHOCULT™ PRODUCTS BE USED FOR LYMPHOID PROGENITOR CFC ASSAYS?
        A. Human lymphoid progenitors (B,NK, and T) seem to require stromal support for growth therefore cannot be grown in MethoCult™. Mouse pre-B clonogenic progenitors can be grown in MethoCult™ M3630.
      • Q. IS IT POSSIBLE TO SET UP CFC ASSAYS IN A 24-WELL PLATE?
        A. Yes, as long as the same ratio of cells is plated. Optimal plating concentration and the number of wells required to get an accurate estimation of CFC numbers may require optimization:
        • The surface area of a 35 mm dish is ~9.5 cm2 and 1.9 cm2 for the 24 well plate.
      • Q. CAN I STAIN THE COLONIES IN THE METHOCULT™?
        A. Colonies can be stained but only if they are plucked from the dish. However, mouse erythroid colonies can be stained in the dish (Benzidine Staining Protocol). CollagenCult™ products are recommended for staining applications. The 3D matrix can be dehydrated and the colonies can then be fixed onto glass slides for immunohistochemical or enzymatic staining.
      • Q. ARE THERE DIFFERENCES IN COLONY MORPHOLOGY WITH SERUM-FREE MEDIA?
        A. The serum containing media generally give better overall growth (especially CFU-GM colonies, as they will contain more cells) but not more colonies. There is no difference in total CFC’s between serum-free and conditioned or recombinant media.
      • Q. CAN METHOCULT™ BE MADE WITH AN ALTERNATE BASE MEDIA?
        A. Yes, this can be done as a 'Custom' media order. Please contact techsupport@stemcell.com for more information.
     

    Data


    Procedure summary for hematopoietic CFC Assays


    Procedure summary for hematopoietic CFC Assays



    Examples of colonies derived from human hematopoietic progenitors


    Examples of colonies derived from human hematopoietic progenitors



    Related Products


    Other Reagents and Materials


    Product Name

    Description

    Catalog #

    Iscove's MDM with 2% FBS Medium for Preparing and Washing Samples for Colony-Forming Cell (CFC) Assays 07700
    35 mm Culture Dishes 35 mm Culture Dishes 27100
    60 mm Gridded Scoring Dishes Dishes for Reproducible and Accurate Scoring of Colonies 27500
    10% Bovine Serum Albumin in Iscove's MDM
    10% Bovine Serum Albumin (BSA) in Iscove's MDM
    09300
    L-Glutamine L-Glutamine (200 mM) 07100
    EasySep™ Human CD34 Positive Selection Kit Immunomagnetic Positive Selection Kit 18056
    EasySep™ Human Whole Blood CD34 Positive Selection Kit Immunomagnetic Positive Selection Kit 18086
    EasySep™ Human Cord Blood CD34 Positive Selection Kit Immunomagnetic Positive Selection Kit 18096
    EasySep™ Human Progenitor Cell Enrichment Kit Immunomagnetic Negative Selection Kit 19056
    RosetteSep™ Human Bone Marrow Progenitor Cell Pre-Enrichment Cocktail Immunodensity Negative Selection Cocktail 15027
    Lymphoprep™ Density Gradient Medium for the Isolation of Mononuclear Cells 07801
    RosetteSep™ Human CD45 Depletion Cocktail Immunodensity Depletion Cocktail 15122
    RosetteSep™ Human Cord Blood Debulking Cocktail Immunodensity Depletion Cocktail 15126
    RosetteSep™ Human Hematopoietic Progenitor Cell Enrichment Cocktail Immunodensity Negative Selection Cocktail 15026
    StemSep™ Anti-Human CD41 TAC Immunomagnetic Selection Reagent 14050
    StemSep™ Human CD34 Positive Selection Cocktail Immunomagnetic Column-Based Positive Selection Kit 14756

    Antibodies


    Product Name

    Description

    Catalog #

    Anti-Mouse CD49f Antibody, Clone GoH3 Rat Monoclonal Antibody Against Human, Mouse, Rhesus CD49f (Integrin α6) 60037

    Cytokines


    Product Name

    Description

    Catalog #

    SCF, Human, Recombinant Stem Cell Factor 02630
    IL-3, Human, Recombinant Interleukin-3 02503
    GM-CSF, Human, Recombinant Granulocyte-Macrophage colony-Stimulating factor (GM-CSF) 02532
    G-CSF, Human, Recombinant Granulocyte Colony-Stimulating Factor (G-CSF) 02615
    IL-6, Human, Recombinant Interleukin-6 02506

    Equipment and Accessories


    Product Name

    Description

    Catalog #

    Blunt-End Needles, 16 Gauge 16 Gauge Blunt-end Needles 28110
    3 cc Syringes 3 cc Syringes 28230
    Ammonium Chloride Solution Reagent for the Lysis of Red Blood Cells 07800

    Publications


    This Product Has Been Used In:


    Background References:


    Recently Viewed Items