Home » Cell Culture Media and Matrix F.A.Q.'s
Cell Culture Media and Matrix F.A.Q.'s
 
  • What are the differences between serum-free, protein-free, and chemically-defined Media? 
     
    Serum-free Media do not contain or require the addition of serum. Animal component-free are Media in which none of the components are animal derived. Protein-free Media do not contain any proteins. Chemically-defined Media contain components which are all known (or defined chemically) and they are usually, but not always, protein-free.
     
  • Media Supplemented with Serum:
     
    Advantages:
     
    1. important source of nutrients, growth factors, hormones, attachment factors, and protection agents
    2. contains trypsin inhibitors
    3. important as a cryoprotectant in cryopreservation
     
    Disadvantages:
     
    1. undefined
    2. lot-to-lot variability, extensive testing required to ensure as much lot consistency as possible
    3. substantial cost
    4. presence of animal-derived components
    5. presents obstacles with purification of products from downstream processing of culture medium
    6. contains growth-inhibitory and growth-promoting activity resulting in an unpredictable inhibition and promotion of Cell growth
    7. standardization of protocols difficult
     
     
    Why should I use serum-free Media?
      
    Serum-free Media offers the customer better lot-to-lot consistency since it contains fewer undefined components, such as serum. The end user also need not take time to qualify lots of FBS for activity. Serum-free Media are lower in protein content than medium supplemented with serum, which can simplify the purification process and increase the yield of the end product.
     
     
  • Media Supplemented with Serum:
      

    Advantages:
     

    1. defined
    2. usually free of animal-derived components
    3. effect of defined components and / or additions to Media on Cell growth may be examined
    4. greater consistency between experiments
    5. purification of desired secreted protein product simplified
     
    Disadvantages:
     
    1. slower Cell growth often observed
    2. must examine the requirements for Cell growth
    3. Cells must be slowly weaned from medium containing serum

 

  • Why should one use undifferentiation Media? 

    The undifferentiating Media is utilized to maintain stem Cells in their undifferentiating state. The undifferentiating Media is to be utilized with the undifferentiating Matrix to maintain the stem Cells. The undifferentiating Media and Matrix allows the stem Cells to express their surface antigens and markers.

     
  • When should one use the expansion Media and Matrix?

    Once the stem Cells are maintained in their undifferentiating Media and Matrix in culture for 7 days, in order to expand the stem Cells one may transfer the Cells to the pre-coated expansion Matrix flasks after subculture. The stem Cells may be expanded in culture in expansion Media and Matrix for 14 days.

     
  • Is there a different Cell freezing Media for the undifferentiating stem Cell condition?

    The Cells when stored in the liquid nitrogen vapor phase in the appropriate Media and stem Cell culturing conditions they are easily recovered whereas when Cells are stored in the liquid phase of the liquid nitrogen if the vials are not properly sealed or if there is a leak the Cells come into direct contact with the liquid nitrogen. Thaw the Cells the viability and recovery of the Cells get comprised in this situation.

     
  • Is there a different Cell freezing Media for stem Cell expansion condition?

    The stem Cells in their expansion condition would require a different set of Cell freezing Media.

     
  • When should one use the Differentiation Media and Matrix?

    After expansion of the stem Cells one may one to differentiation the stem Cells into an appropriate Cell type and depending on the Cell type one would utilize the appropriate differentiation Media and Matrix combination. Usually it should take anywhere from 7 – 14 days in differentiation Media and Matrix stem Cell culturing conditions to obtain the desired end results. During this time frame in the differentiation culture conditions one may subculture the Cells in the differentiation Media and Matrix environment.

     
  • Is there a different Cell freezing Media for stem Cell Differentiation condition?

    The stem Cells in their differentiation condition would require a different set of Cell freezing Media. Freezing the Cells in their differentiation Media and Matrix environment will enable one to go back a re-visit their results at that stage of the stem Cell differentiation process.

     
  • Why is there a different Media and Matrix for each stage of stem Cell culturing conditions?

    The various stages of stem Cell growth and proliferation require a unique niche and in tissue culture we are able to manipulate the Media and extracellular microenvironment of the stem Cells to optimize their growth and proliferation at the various stages (undifferentiation, expansion and differentiation). 
    The expansion media and matrix maintains the cells in their proliferation stage of growth. The difference between expansion media and growth media is that the expansion media and matrix has a faster doubling time then when cells are cultured in growth media and matrix. The growth medium and matrix the cells are maintained through out the spectrum of the cell cycle. The doubling time of the cells in the growth media is slower when compared with the expansion media and matrix. The undifferentiation medium and matrix has a slower doubling time when compared to the growth media and expansion media. The purpose of the undifferentiation medium and matrix is for maintaining the cells in their undifferentiation stage of the cell cycle and allows the expression of the biological markers within their niche. Initially, when one thaws the cells we recommend that they utilize the growth media and matrix as indicated in the cell culture flow diagram.