I have to dissociate rat (7days old) testis for obtaining Sertoli cells. I have the protocol and I have to use a solution of Collagenase IV and DNase I (M0303S BioLabs) in DMEM.
My problem is that in the protocol I have the DNase concentration in mg (10mg/ml) but in the data sheet of DNase I there is the concentration in units.
Usually I have to dissociate 10 testis in 20ml digestion solution.
Can someone suggest which is the DNase I (BioLabs) concentration in units I have to use?
Hello alessandra, for obtaining Sc from 7d old rat testis use of DNase not necessary, however if needed you can use 50units of DNase I which shoud be good enough.
I haven't isolated sertoli cells from testis before, but I use a combination of Liberase TM and DNAse I, grade II (Roche Molecular) to isolate lymphocytes from murine GI and female reproductive tract. I have found the DNase is helpful when you are trying to gain single cell suspensions as the DNA from dead/dying cells can inhibit the isolation process. If you are interested in switching the DNase you use, the DNase we use comes in 100mg aliquots and is 2,000 Kunitz units per mg; I use 100ug/ml (final concentration). You could also probably call the company and ask if they have the conversion for you. Good luck.
I'm going to use DNAse I and liberase to isolate ovarian follicles. I red that dissociation can be done in DPBS but at the same time the DNAse I should have optimal activity at 5mM Mg2+, i.e. an Mg concentration ten fold higher than in DPBS. Do you know other suitable buffers for tissue dissociation with DNAse I?
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