Cat: MHF1
Cat: MHF1
SIM HF Expression Medium is an optimized serum-free medium used to support the growth of High FiveTMcell.It is for research use only not for use in human or animals. This medium contains L-glutamine and is ready-to-use. |
• Specifically optimized for the culture and transient transfection of Hi5 cells |
• Strict quality control, stable performance, cost-effective |
• Serum-free, protein-free |
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• Media: SIM HF Expression Medium |
• Cell Line: BTI-TN-5B1-4 (Hi5) cells |
• Culture Type: Suspension |
• Culture Vessel: Shake flask, bioreactor, etc. |
• Temperature Range: 26℃ to 28℃ |
1. Rapidly thaw (less than 2 minute) avail of frozen Hi5cells in a 27℃ water bath; 2. Decontaminate the vial and transfer the thawed cells into a sterile centrifuge tube with 5-10 mL fresh medium; 3. Centrifuge the cells at 200 × g for 5 minutes, remove the supernatant slowly. 4. Suspend the cells with fresh medium in sterile shake flask, and culture in an orbital shaker at 150 rpm; 5. Subculture the cells when the cell density is ≥2-3× 106 viable cells/mL. |
1. Seed cells into pre-warmed fresh SIM HF Expression Medium at a final density of 0.4-0.5×106 viable cells/mL. Incubate the cells at 27±1℃ rotating at 150 rpm(The speed may vary among different shakers. Resuscitated cells are recommended for adaptive culture at a lower rotational speed); 2. Subculture the cells again, once cell density reaches 2-3 ×106 viable cells/mL. |
1. Determine the density of cells in logarithmic phase with a viability ≥ 90%; 2. Prepare cell freezing medium which is composed of 45% fresh SIM HF Expression Medium, 45% conditioned medium(cell culture supernatant of the cells used for cryopreservation)and 10% DMSO; 3. Centrifuge (200 × g, 5minutes) and remove all the supernatant. Resuspend the cells with suitable volume of cell freezing medium to viable cell density; 4. Aliquot the cells to cryovials and place to controlled-rate freezing apparatus. Put the apparatus in -80℃ refrigerator; 5. Transfer the tubes to liquid nitrogen for long-term storage. |
(1).Direct adaptation |
In general, Hi5 cells can be directly adapted from serum-reduced media or other serum-free media into SIM HF Expression Medium. Cells used for adaptation is required to be in the logarithmic phase with a viability ≥ 90%. 1. Subculture the cells directly into 20 mL of fresh SIM HF Expression Medium at a final density of 0.5-0.7×106 viable cells/mL; 2. Incubate the cells at 27℃, rotating at 150 rpm; 3. 3-4 days later, determine the cell density and viability. Generally, the cell density should be > 2×106 cells/mL, which means that direct adaptation is successful. |
(2). Sequential adaptation |
The cell inoculation density was also 0.5-0.7×106 cells/mL. In the passage process, the ratio between the primary used medium and SIM HF Expression Medium in the mixed medium gradually decreased from 90:10 to 75:25, 50:50, 25:75, 10:90, 0:100. Note: In the process of cell adaptation, the first three generations of cells may not be good, but generally the cell state will improve from the fourth generation. |