IM-T T-lymphocyte culture medium
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)
Day5 Cells are transferred to gas-permeable bags. A total of 1,000 mL
Day8 The number of gas-permeable bags is increased. A total of 2,000 mL
Day14 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
iMediam for T
Human T-lymphocyte culture
|Tリンパ球培養用基礎培地||RPMI-1640+7s||(IL-2 free, 血清free)|
|Tリンパ球培養用培地||RPMI-1640+9s Sfree||(IL-2 700U/ml, 血清free)||初期培養にお勧めです|
|Tリンパ球培養用バッグ培地||CP-4(LL7.3) Sfree||(IL-2 350U/ml, 血清free)||増幅培養にお勧めです|