Top : Cell Biology : Cell Culture : General Procedures : Maintenance of Cell Line
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Protocols
  • Subculture of Adherent Cell Lines (Sigma)
    Adherent cell lines will grow in vitro until they have covered the surface area available or the medium is depleted of nutrients. At this point the cell lines should be sub-cultured in order to prevent the culture dying. To subculture the cells they need to be brought into suspension. The degree of adhesion varies from cell line to cell line but in the majority of cases proteases, e.g. trypsin, are used to release the cells from the flask...
    http://www.sigmaaldrich.com/life-science/cell-cult...
    Added: Fri Feb 06 2009, Hits: 3305, Reviews: 0  Write review   Cached
  • Lymphoblastoid Cell Lines from Frozen Whole Blood (Donis-Keller lab)
    Blood Samples can be stored frozen as a backup in case an LCL is needed at a later date.
    http://wheat.pw.usda.gov/~lazo/methods/donis/hcc/h...
    Added: Tue May 14 2002, Hits: 232, Reviews: 0  Write review   Cached
  • Lymphocyte Transformation (Donis-Keller lab)
    Lymphocytes are transformed to establish cell lines. Mononuclear cells (lymphocytes) from anticoagulated venous blood are isolated by layering onto histopaque. During centrifugation, erythrocytes and granulocytes are aggregated by ficoll and rapidly settle to the bottom of the tube; lymphocytes and other mononuclear cells remain at the plasma-histopaque interface. Erythrocyte contamination is neglible. Most extraneous platelets are removed by low speed centrifugation during the washing steps.
    http://wheat.pw.usda.gov/~lazo/methods/donis/hcc/h...
    Added: Tue May 14 2002, Hits: 572, Reviews: 0  Write review   Cached
  • Maintenance of Cell Culture ( Contributed by Nanci Donacki )
    Detailed procedure for culture and subculture cell in flask and plates
    Added: Tue May 14 2002, Reviews: 0  Write review  
  • Preparation of Lymphoblastoid Cell Lines for Long-Term Storage (Donis-Keller lab)
    To store cell lines in a form that will insure recovery with high viability. A culture in logarithmic phase of growth with a total volume of 80-100 ml/T-75 flask should yield enough cells to freeze 10 ampules (1.0 ml/ampule). Cells should have a count of 4 X 106 cells/ampule to 9 X 106 cells/ampule. Too high or too low a cell count lowers recovery viability. Cell are frozen in RPMI-1640 with 15% Fetal Bovine Serum + 10% DMSO. Cultures are frozen slowly using a Model 700 Controller freezing chamber. This precision electronic device automatically controls the injection of liquid nitrogen into the freezing chamber to provide a 1 degrees C/minute freezing rate from +4 degrees C to -45 degrees C (with automatic heat of fusion compensation), then a 10 degrees C per minute freezing rate to -90 degrees C. Frozen ampules should be stored in liquid nitrogen for long term storage or in a -135 degrees C Cryopreservation System. Note: Cryotubes should be labeled with cell line number anddate prior to beginning this procedure.
    http://wheat.pw.usda.gov/~lazo/methods/donis/hcc/h...
    Added: Tue May 14 2002, Hits: 168, Reviews: 0  Write review   Cached
  • Subculturing Cells (Yu-li Wang's Lab, University of Massachusetts Medical School )
    Basic procedures for routine cell culture
    http://ylwang.umassmed.edu/protocol/cc/subcultu.ht...
    Added: Fri Oct 17 2003, Hits: 1713, Reviews: 0  Write review   Cached
  • Subculturing Monolayer Cell Cultures PDF file (Corning)
    This 5-page protocol provides a detailed procedure for subculturing attached cells using enzymatic dissociation. It explains the theory behind some of the keys steps in the process. This link also works:http://catalog2.corning.com/Lifesciences/media/pdf/lp_cellculture_selection_guide_3_02_lslp_cc_006.pdf
    http://www.fisher.co.uk/techzone/pdfs/Subculturing...
    Added: Wed Feb 11 2009, Hits: 1350, Reviews: 0  Write review