I apologize my ignorance! What is WAT? We are working with murines like rats and mouse usually. We used from 550±250 mg from mouse epididymal fat and 1-2g from rat epididymal fat.
Hi, thanks for the answer! WAT means White Adipose Tissue. So you do not use inguinal fat? How many vital cells can you obtain from 550 mg?
Thanks
I believe that we are working with the same fat type, with different semantic words, why the epididymal fat is in the inguinal region! 𝑥̅ =6,44×106±7,44×106 from mouse and x108 from rat!
The MSCs from fat always have a good yield!
Alright! I call the subcutaneous "inguinal" and the visceral "epididymal".
Anyway, thank you very much! Actually I have obtained very few cells from one mouse 7 weeks old. Can I ask you to share the isolation protocol you are using?
Obrigado!
Regarding the semantics: the inguinal fat is quite easily distuingishable from epididymal fat, as the inguinal fat is more superiorly and laterally located, while the epididymal fat is more inferiorly and medially located in mice. The consistency and colour is also slightly different. Both are considered white adipose tissue.
Hello Yuri, I remembered this discussion and now I have done this multiple times, I can give a more reliable answer. The average yield of stromal vascular fraction cells was 1.98 x 10^6 per mouse. The average yield of inguinal adipose tissue (after removing the lymph node) was 0.18 gram per mouse. So, on average 1.1 x 10^7 cells per gram of adipose tissue. The mice were male 7 to 9 weeks old. The cell viability was >90%.
Getting 70% confluency at P0 took me always a bit longer (around 1 week at initial plating density of 1x10^6 SVF-cells per 100-mm dish, with a first medium change after 24 hours) than getting 70% confluency at P1, P2, P3,... (2-3 days at plating density of 2x10^5 cells per 100-mm dish). Good morphology and cell surface marker expression (CD44+, CD73+, CD90.2+, CD105+, CD11b-, CD31-, CD45- and CD34-).
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