■ Comparison of the performance of a medium containing serum, which is currently used at our company, with “iMediam for T” (Day 14 of culture)
Peripheral blood mononuclear cells (PBMC) were separated from the blood of 9 healthy individuals who gave informed consent and cultured using our medium containing serum and “iMediam for T” containing no serum to compare the cultures. The cells were cultured in OKT3 antibody-immobilized flasks until day 5 of the culture, and then were transferred to gas-permeable bags and cultured for 14 days in total. The final liquid volume was 2,000 mL.
Test protocol: iMediam for T
Container: OKT3 antibody-immobilized flask 225 Cells: PBMC from healthy individuals; 2 × 107 cells/flask (50 ml IL-2: 700 U/mL)
Cells are transferred to gas-permeable bags. A total of 1,000 mL
The number of gas-permeable bags is increased. A total of 2,000 mL
Cell number measurement, surface antigen analysis
■ Surface antigen analysis
Peripheral blood mononuclear cells (PBMC) from 3 healthy individuals who gave informed consent were cultured using “iMediam for T” containing no serum for 14 days. Then, the ratio of CD3-positive cells, which is a T-lymphocyte index, was measured by a flow cytometer. The ratio of CD3 was 98% or above in all 3 specimens, which indicates that T-lymphocytes were selectively propagated.
- * Application: Human T-lymphocyte. * Storage condition: Cold dark place (2–10ºC).
- * Sterility tests: ○ Endotoxin: Colorimetric method ○ Mycoplasma: Culture method ○ Fungus and bacteria: Agar plate surface-spread method
- * Procedure: (1) Prepare OKT3 antibody-immobilized flask 225. (2) Suspend 3–5 × 107 cells of PBMC into 30–50 mL of “iMediam for T”. (3) Add an appropriate amount of “iMediam for T” depending on the condition of cell proliferation. (4) After 5–7 days of culture, transfer the cell suspension to gas-permeable culture bags, add an appropriate amount of “iMediam for T”, and continue to culture for another 2–3 weeks. Increase the number of culture bags depending on the culture condition.
- * Notes: ○ Do not use on the human body. ○ We shall not be held responsible for accidents or damage resulting from using this product. ○ If you have any questions about the usage, please contact GC LYMPHOTEC, Inc.
|Product code||Product name||Use||Volume|
|iM-T||iMediam for T||Human T-lymphocyte culture||1,000mL|