Hello,
I am trying to transduce my CD4/CD8 T-cells with lentivector using retronectin from Takara (#T100B) at 50ug/ml, after they are activated by CD3/CD28 for 2 days.
The official protocol says that "it's important that the target cells be in logarithmic growth phase", but how this state may be monitored?
How to choose the "perfect moment" for transduction via retronectin?
Thanks in advance!
Hi, Mark
I suggest that in a few samples, the greater the number the better, you should do a total cell count at different times and plot the number against time. From this you should be able to identify the initial lag phase, the subsequent log phase and even the final plateau phase. From this you should be able to identify the most logarithmic growth period
Narayanan
I agree. You have to count your cells at the begining of the replication, and at different times since then. For example, at 24, 48 and 72 h. The time you found the biggest increase of the number of cells in respect those you had the previous time, is the log phase time.
For example, if you counted:
0 h: 100,000
24 h: 250,000
48 h: 1,000,000
72 h 1,200,000
The log phase of your cells is 48 h.
I hope you have understood,
good luck!
- Badges
- Science topic
- Similar topics
- Instrumentation Engineering
- Instruments