ClonaCell™ FLEX

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Base Methylcellulose Medium for the Selection and Cloning of Cells

  • ClonaCell™ FLEX 45 mL
  • Label for ClonaCell™ FLEX Medium 45 mL
ClonaCell™ FLEX 45 mL
ClonaCell™ FLEX is a base methylcellulose medium that is combined with a 2X concentrate of the user’s own liquid medium to create a custom semi-solid cloning medium. This allows the user to adapt an existing liquid cloning method to a customized semi-solid cloning method. ClonaCell™ FLEX is recommended for selection and cloning of suspension-adapted mammalian cells, including CHO cells and hybridomas. The medium is chemically defined, protein-free and animal component-free. It does not contain L-glutamine or selection agents. The medium is compatible with a wide range of selection systems, including dihydrofolate reductase (DHFR), glutamine synthase (GS) and hypoxanthine-aminopterin-thymidine (HAT).
 
Benefits of semi-solid cloning:
• During semi-solid cloning, individual cells are suspended in viscous medium, which physically separates them and allows them to grow into discrete colonies that can be easily isolated.
• With semi-solid cloning, monoclonal cell lines can be isolated in less time and using fewer resources than with limiting dilution cloning.
• It is easy to isolate rare, high-producing clones, compared with selection in bulk liquid medium. This is because diverse clones with a wide range of growth rates and productivities form discrete colonies.
Product Name Description Catalog # Size Price Quantity
ClonaCell™ FLEX Base methylcellulose medium for the selection and cloning of cells 03818 45 mL 227.00 USD      
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Application: Cell line development, Custom media, Hybridoma generation
Product Type: Hybridoma generation & transfected cell selection
Recommended For:
• Creating a customized semi-solid medium for selection and cloning of suspension-adapted mammalian cells, including CHO cells and hybridomas
• Converting a user-selected liquid medium into a semi-solid medium for selection and cloning of a wide variety of cell types
• Recloning of established suspension-adapted mammalian cell lines
Accessory Products:
Blunt-end Needles (Catalog #28110/28120)
Contains:
Methylcellulose concentrate
Advantages:
• Allows the user to convert an existing liquid cloning method into a semi-solid cloning method using a 2X concentrate of their own liquid medium.
• The ability to create a custom methylcellulose-based semi-solid medium allows semi-solid cloning of a wide variety of mammalian or non-mammalian cell lines that can be cultured in liquid medium.
Area of Interest: Cell line development, Embryonic stem cell & iPS cell research, Hematopoietic stem cell research, Hybridoma generation
Cell Type: CHO, Embryonic stem cells & iPS cells, Hematopoietic stem & progenitor cells, Hybridomas
Medium Type: Methylcellulose-based, Serum-free
Popular Product Line: ClonaCell™
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.

Protocols and Manuals

Current documents
Product NameLot# ?Document TypeDownload
ClonaCell™ FLEX All Manual
ClonaCell™ FLEX All Product Information Sheet
 

Educational resources:

MSDS:

FAQS:


    • Q. WHY DO I GET MORE CELLS WHEN I SELECT MY FUSION IN LIQUID MEDIUM RATHER THAN IN METHYLCELLULOSE-BASED SEMI-SOLID MEDIUM?
      Cells grown in liquid medium may appear to grow more rapidly than in methylcellulose-based medium. This is often due to the presence of a few rapidly growing clones that multiply quickly and become abundant in liquid culture, overgrowing clones that grow more slowly. In methylcellulose cultures, the rapidly growing cells remain in close proximity to each other, resulting in large colonies, each derived from a single fusion or transfection product. The large clones don't overgrow smaller, slower growing colonies, which can be separately isolated.
    • Q. HOW DO I THAW CLONACELL™ METHYLCELLULOSE-BASED SEMI-SOLID MEDIUM?
      We recommend thawing the medium overnight in a refrigerator at 4°C and mixing well.
    • Q. HOW DO I MEASURE AND DISPENSE METHYLCELLULOSE SEMI-SOLID MEDIUM?
      We recommend using a 12 mL syringe with a 16 gauge needle attached (blunt end needles are recommended for safety purposes). Do not dispense the semi-solid media/cell mixture using serological pipettes as the media will stick to the pipette walls, resulting in inaccurate dispensed volumes and loss of cells.
    • Q. MY CLONACELL™ METHYLCELLULOSE SEMI-SOLID MEDIUM APPEARS RUNNY. WHY DOES THIS HAPPEN?
      "Runny" methylcellulose could be a result of improper handling. Diluting the methylcellulose with too much liquid medium, or insufficient mixing before use, will result in methylcellulose with altered viscosity. Excessive condensation on the inside of the cell culture dish lid can result in water dripping onto the cultures, lowering viscosity. Additionally, bumping, shaking or other sudden movement of the culture may also disrupt the colonies. Note: methylcellulose is less viscous at room temperature than at 37°C.
    • Q. WHAT IS THE OPTIMAL NUMBER OF COLONIES PER PLATE?
      We recommend 50-150 colonies per plate. As it is difficult to anticipate the numbers of colonies after a fusion or transfection, we recommend plating at three different densities to increase the likelihood of achieving a plating density of approximately 100 colonies per plate. This density allows sufficient space between the colonies to allow for easy colony picking.
    • Q. THERE ARE STILL BUBBLES IN THE MEDIA AFTER I PLATE MY CELLS. DO I NEED TO DISRUPT THE BUBBLES?
      We recommend that you avoid creating large bubbles during plating, but there is no need to manually pop or disperse the small bubbles after plating. They will disperse over the incubation period of 10-14 days.
    • Q. DO I EVER NEED TO RE-CLONE CULTURES GROWN WITH CLONACELL™ SEMI-SOLID MEDIUM?
      Re-cloning is a good practice to observe and is recommended if the number of colonies in the original dishes was very high.
    • Q. ONCE I PICK THE COLONIES AND GROW THE CELLS IN PLATES, WILL THE RESIDUAL METHYLCELLULOSE INTERFERE WITH CHARACTERIZATION? FOR EXAMPLE, WILL I HAVE PROBLEMS DOING AN ELISA?
      There will likely be some residual methylcellulose contamination when colonies are picked and transferred to the 96-well plate with the liquid growth medium. The concentration of methylcellulose, however, should be low enough that it should not interfere with most assays.
    • Q. HOW IMPORTANT IS THE INCUBATOR HUMIDITY WHEN CULTURING IN METHYLCELLULOSE-BASED MEDIUM?
      A. Very important. In situations where the humidity is not high enough, we recommend that the 100 mm Petri dishes should be placed with an open dish containing sterile water inside a larger plastic container with a lid. Without very high humidity, the media will dry out over the culture period and this will impede the growth of the colonies.
    • Q. DO I HAVE TO USE 100 mm PETRI DISHES OR CAN I USE OTHER CULTUREWARE?
      We recommend 100 mm Petri dishes as these have been used to develop and test ClonaCell™ semi-solid media. We have found that the surface area of these dishes allows for easy colony picking. Other sizes of dish (e.g. 6-well plates) can be used. It is important to use non-coated dishes to prevent cells from sticking to the bottom of the plate and obscuring the colonies. The volume of media plated should be adjusted to reflect the surface area of the dish being used.
 

Product Name

Description

Catalog #

ClonaCell™-CHO CD Medium Transfected CHO Cell Selection and Cloning Medium 03815
ClonaCell™-HY PEG Polyethylene Glycol for Cell Fusion 03806
100 mm Treated Tissue Culture Dishes 100 mm Treated Tissue Culture Dishes 27125
ClonaCell™-CHO ACF Medium Transfected CHO Cell Selection and Cloning Medium 03816
ClonaCell-CHO CD Liquid Medium Chemically-Defined, Serum-Free, Protein-Free, Animal Component-Free Liquid Medium for Culture of CHO Cells 03817
ClonaCell™-TCS Medium Transfected Cell Selection and Cloning Medium 03814
ClonaCell™-HY Medium A Pre-fusion Myeloma Growth Medium and Hybridoma Expansion Medium 03801
ClonaCell™-HY Medium C Fusion Recovery Medium 03803
ClonaCell™-HY Hybridoma Kit Complete Kit for Mouse Monoclonal Antibody Generation 03800
ClonaCell™-HY Medium E Hybridoma Growth Medium Containing HT 03805
Blunt-End Needles, 16 Gauge 16 Gauge Blunt-end Needles 28110
ClonaCell™ EasyPick Robotic Colony Picking and Liquid Handling Instrument 30000
ClonaCell™-HY Medium D Methylcellulose-Based Hybridoma Selection and Cloning Medium 03804
ClonaCell™-HY Medium B Hybridoma Fusion Medium 03802

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