I am working with Balb/c mice, and in my experiment, I have to inject mice with antigens, then get their spleen and collect the splenocytes for fusion. Recently, I get into trouble with the mouse spleen: it's much smaller than normal and of course, it leads to a small number of splenocytes.
This is my experiment: I had 5 mice, all these 5 mice were injected with the same protocol (1 type of antigen, the same doses, injection route; the same nutritional and habitat conditions). after immunization, I collected the spleens and obtained the splenocytes from 5 mice (cut the spleen into pieces, crushed them by the syringe plunger flange, then filtered the splenocytes through a cell strainer 40um), the numbers of splenocytes were collected as follows:
- Mouse 1: 4x10^7 cells
- Mouse 2: 1x10^8 cells
- Mouse 3: 1.5x10^8 cells
- Mouse 4: 6.7x10^7 cells
- Mouse 5: 2.3x10^7 cells (the smallest spleen)
Does anyone be in this situation? Please share with me the causes and how can I overcome this issue?
Many thanks!
Hi Maria Valsjö,
I did not measure them, but they were smaller than the previous ones I've done (I've done the experiment with over 50 mice before and "the normal" means I compared with those 50 spleens).
And based on the cell counting results, I usually collect 10^8 - 2x10^8 splenocytes per spleen, but this time, the number of cells were much more less than before (as I mentioned above), using the same technique for collecting splenocytes.
And we used the same source of Balb/c (the same provider), had no big different in breeding condition.
Maria Valsjö, could you share with me how many splenocytes you've collected from a immunized-mouse spleen?
Dear Thao, I have not collected splenocytes but I've worked with Balb/c mice for over 20 years so I that's why I wondered about the size. I know the size sometimes is different between litter batches but its not usually a huge different. Did you give them the same concentration?
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