hello everyone,
I have a question about my experiment related to preadipocytes differentiation. I am using subcutaneous preadipocytes of cattle, when I passage them in 6 well plates and after 80-90% confluency wash with PBS and add differentiation medium (2% horse serum) and see cell under microscope many cells are washed off, and even during differentiation either i wash cells with PBS or change medium with out wash still some cell aspirate with medium and after adding new medium many cells start floating in the medium, i tried many times and change medium very carefully.
this problem even more severe in 12 well plate
i also found some cell looks strange
kindly answer if anybody know about these issues...
some pictures attached
1. preadipocyte confluency must be 100% before adding differentiation media.
2. serum percentage will be 10% in Differentiation media.
for chemically induced adipogenesis ATCC prescribed method can be followed.
Link:
https://www.atcc.org/resources/technical-documents/chemically-induced-method-of-adipogenesis
Debarun Patra thanks for your response. in our lab we mostly use 10%FBS for cell proliferation and 2-5% Horse serum for differentiation with any respective treatment.
Hello Sajida Naseem
I am unaware of the differentiation of cattle preadipocytes, but I have differentiated the mouse and human cells into adipocytes.
I want to suggest some of the points which might be helpful for you.
I hope these points will help you to let them grow and differentiate into healthy adipocytes.
All the best 👍
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