Culture of oct-4 positive cells from preputial epidermis

The medical waste of preputial skin is easily obtained in Indonesia. The cells isolated from six samples are expected to have pluripotency and able to differentiate to other lineage for medical treatment. This research uses the epidermal layer of preputial skin to obtain keratinocytes, this cells are taken using dispase and trypsin solution overnight respectively. Then, the keratinocytes are subsequently cultured using DMEM complete high glucose to increase the number of cells. The cultured cells are then taken for immunocytochemistry (ICC) of Oct-4 since it is the marker of pluripotency. The other half of cultured samples are continued for over confluency analysis for fourteen days to observe spontaneous differentiation. This research has successfully used an alternative and cost effective culture medium for keratinocytes. It consist of DMEM high glucose, PRP 10%, heparin 1%, FBS 10%, penstrep 1%, and fungizone 1%. The result of ICC is partially positive with keratinocytes nuclear being stained dark brown in five hpf from each sample. However, spontaneous differentiation analysis using alcian blue shows negative result of chondrogenic formation from keratinocytes.