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    Home > Chemicals Industry > Chemical Technology > Mammalian cell culture medium (3)

    Mammalian cell culture medium (3)

    • Last Update: 2021-10-02
    • Source: Internet
    • Author: User
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    4 Cultivate for 48h~72h, observe the cell morphology, and count according to 7.
    7.
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    3
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    5 When the cell density reaches 1×10 5 cells/mL, replace 10 mL of cell culture medium without calf serum , continue culturing for 48 hours, observe the cell morphology and count
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    4.
    3 Cell count

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    1 Prepare cell suspension B according to the method of 7.
    7.
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    1 for the cells to be counted
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    2 Pipette 100uL of cell suspension B and transfer it to a centrifuge tube, and determine whether to dilute and the dilution factor depending on the amount of cells
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    3 Place a cover glass over the counting chamber in the center of the counting plate
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    Adjust the number of cells in the counting plate to 100-300
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    Add the cell suspension along the edge of the cover glass to make it naturally infiltrate through capillary action.
    Do not leave bubbles or overflow, and count under the microscope
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    4 Observe the number of cells in the large squares at the four corners of the counting plate.
    When the cells are pressed to the midline, generally follow the principle of "counting left but not right, counting up but not down"
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    Substitute the number of cells observed on the counter into the following formula:

    The number of cells observed on the counting plate × 10 4 × the dilution factor/4 = the number of cells/mL

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    5 Expression of analysis results

    Cultured cell morphology 48h ~ 72h for fibroblast-like, adherent growth, normal cell morphology see Appendix A
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    The number of cells should reach 1×10 5 cells/mL within 72 hours
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    48h cultured cell morphology of fibroblast-like, adherent growth, normal cell morphology see Appendix A
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    The cell count should not be less than 1×10 5 cells/mL
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    8 Inspection rules

    8.
    1 All items in the technical requirements of Table 2 are factory inspection items
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    2 The homogeneous products produced by the same mixing equipment at one time are one batch
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    Each batch must be inspected
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    3 Sampling according to the provisions of GB/T6679-2003
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    Divide the sampled products into two evenly, and put them in two clean and dry sterile, airtight, light-proof containers, and paste the labels to indicate the name of the manufacturer, product name, batch number, production date, sampling date and sampler’s name
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    One is for inspection, and the other is kept as a reserve sample for future reference
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    8.
    4 The product should be inspected by the quality inspection department of the manufacturer
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    The manufacturer shall ensure that the products delivered out of the factory meet the requirements of this standard, and each batch of products delivered shall be accompanied by a quality certificate in a certain format, which includes: the name and address of the manufacturer, product name, batch number, date of production and serial number of this standard.
    and so on
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    5 The inspection result is judged according to the GB/T1250 rounding value comparison method.
    If an indicator of the inspection result does not meet the requirements of this standard, it shall be sampled from twice the amount of packaging unit for re-inspection, even if the re-inspection result is only one If any index does not meet the requirements of this standard, the entire batch of products is unqualified
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    9 Labels, instructions for use

    9.
    1 Label

    There should be product name, subtype code, batch number, packaging specification, production date, expiration date, manufacturer name, preparation method, number of this standard, and labelled quantity per liter
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    9.
    2 Manual

    There should be product name, subtype code, packaging specifications, expiration date, manufacturer name, preparation method, usage method, and labelled quantity per liter
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    10 Marking, packaging, transportation and storage

    10.
    1 Logo

    The packaging container should have the product name, subtype code, batch number, production date, expiration date, packaging specification, quantity, number of this standard, etc.
    and the "upward" and "stacking layer limit" signs specified in GB/T191-2000.
    Graphic signs
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    2 Packaging

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    1 Inner packaging

    The materials should meet the national drug packaging material standards, have no precipitates, do not react with mammalian cell culture media, can achieve sealing and light-proof effects, and should be sterilized by irradiation
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    2.
    2 Outer packaging

    It has a light-proof effect, free fall one meter from the ground, and the packaging should not be damaged
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    3 Transportation

    Use enclosed means of transportation
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    The means of transportation should be clean, hygienic, and dry, and must not be mixed with toxic or unclean materials for shipment
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    Avoid sunlight, rain, moisture and pollution during transportation
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    4 Storage

    The product should be stored in a warehouse at 2℃~8℃
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    The warehouse environment should be clean
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    The packaging of the product can be stacked logo, but it should be regular maintenance, such as turning the stack, visual inspection, sampling and so on
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    Under the above storage conditions, the product is valid for two years from the date of production
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