Extraction of Primary Human Keratinocytes and Fibroblasts from Adult Foreskin
Main Article Content
Abstract
Currently, the extraction of skin keratinocytes and fibroblasts for 2D and 3D cultures is an important tool for research in dermatology, including studies of activity and toxicity properties of agents that are in the process of skin product development. The objective of this study was to extract keratinocytes and fibroblasts from adult foreskin for research purpose. The foreskin samples were obtained from volunteer subjects who underwent foreskin surgery at Lerdsin Hospital. Following DPBS washing step, the skin samples were trimmed off fat and connective tissues. The foreskin was cut into small pieces and digested with dispase. The epidermis and dermis layers were separated and the epidermis layers were then trypsinized to obtain keratinocytes while the dermis was placed onto culture dish to extract fibroblasts. The keratinocytes in passage 1 and 2 grew normally in specific keratinocyte culture media. However, cells at passage 3 showed upward proliferation; thus, prohibiting further culture to increase cell number. On the other hand, fibroblasts proliferated well in normal culture media. In summary, the extraction of keratinocytes from adult foreskin is suitable for short-term cell culture experiment. If higher cell number is desired, pooled samples should be performed in order to obtain adequate cells for 3D skin model construction. In contrast, the extraction and cultivation of primary fibroblasts were easy for general cell culture laboratory.
Article Details
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