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  4. GC Lymphotec iMediam™ for T
Serum-free medium

GC Lymphotec iMediam™ for T

iMediam for T is a medium for culturing human T lymphocytes.
iMediam for T allows T cell to proliferate more efficiently than culture using anti-CD3 antibody alone by activating T cell with anti-CD3 antibody/IL-2.

Features

  • Allows serum-free culture of T cells
  • Contains no heterogeneous components (Chemically-defined medium)
  • Obtained the Confirmation of Product Material Eligibility of Regeneration Medicines etc.
    (Notification No. 0110012 issued by the Director of the Center for Product Evaluation, Pharmaceuticals and Medical Devices Agency)
Application :

Human T-lymphocytes

Storage conditions :

Cold dark place (2-10 ℃)

Sterility tests :
  • Endotoxin : Colorimetric method
  • Mycoplasma : Culture method
  • fungus and bacteria : Agar plate surface-spread method

Procedure

  1. Dilute OKT3 antibody with PBS to 5–10 μg/mL, and spread on the bottom of the flasks. (The concentration when using a 225 cm2 flask is 10 µg/mL.)
  2. Suspend PBMC of 3–5 × 107 cells into 30–50 mL iMediam for T.
  3. Add appropriate amount of iMediam for T depending on the condition of the cell growth.
  4. After 5 to 7 days of culture, transfer the cell suspension into a gas-permeable culture bag. Add appropriate amount of iMediam for T, and continue culturing for additional 2 to 3 weeks. Increase the number of culture bags depending on the condition of the culture in an appropriate manner.

Example of Use

Comparison of the peformance of a medium containing serum, which is currently use at GC LYMPHOTEC, with "iMediam for T" (Day 14 of culture)


  • GC Lymphotec isolated PMBC from the blood of nine healthy people who had given informed consent, and compared the culture results using serum-containing culture medium and iMediam for T (serum-free). We cultured the cells in OKT3 antibody-coated flasks for up to five days, and then transferred them to gas-permeable culture bags, where they were cultured for a total of 14 days. The final volume of the culture medium was 2,000 mL. iMediam for T produced results equivalent to or better than those obtained with serum-containing culture media.

  • Protocol

    Container: OKT3 antibody personalized flask 225
    Cells: Healthy donor PMBC 2 x 107 cells/Flask
    (50 mL IL-2:700 U/mL)
    Day 5
    Transferring cells to a gas-permeable culture bag
    (total volume: 1,000 mL)
    Day 8
    Increase the number of cells in the gas-permeable culture bag
    (Total volume: 2,000 mL)
    Day 14
    Cell Counting, Surface Antigen Analysis

Surface Antigen Analysis


After culturing PMBC from three healthy individuals who had given informed consent for 14 days in iMediam for T (serum-free), the percentage of CD3-positive cells, which is an indicator of T lymphocytes, was measured using a flow cytometer. The percentage of CD3 in all three samples was 98% or more, indicating that T lymphocytes were selectively proliferating.

Electron microscope/Giemsa stain

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For research use or further manufacturing use only. Not for use in diagnostic procedures.

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