I've tried different ways of cutting it into small pieces but no matter how much I cut, they still look quite big. I think this is a reason why I cannot get higher cell numbers in my single cell suspension.
I will be very grateful for some practical advice.
I process human tissue to get primary skin cells for cell culture. After disinfecting the tissue, I wash it three times in PBS. Then I trim off as much of the subcutaneous fat layer, as is possible. Afterwards, I cut up the remaining tissue (very small pieces) into a small dish of trypsin (you can also use dispase) that I cover and store at 4C O/N. The next day, I place the dish at 37C for 5-10 min and then I strip off the epidermis using 2 forceps. The epidermis is placed into a conical containing appropriate medium that will support the growth of my cell type desired (FB, MC or KC). I vortex the tube twice, 1 min each, on the highest speed to help break up the tissue. Then I transfer all medium/epidermal tissue into an appropriate flask (T25 or T75). You can add extra medium to help support your cell growth.
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