The Human Skin-Derived Precursors for Regenerative Medicine: Current State, Challenges, and Perspectives

Figure 2

The protocols for hSKPs culture. (a) Traditional method to culture hSKPs: (I) skin samples are cut into pieces, and the epidermis is removed after enzymatic digestion. (II) The pieces are mechanically dissociated into single cells. These two steps are known as cell isolation, and they usually take 1-2 days for isolation procedure. (III) Single cells are seeded into SKPs medium containing bFGF, EGF, and B27, and primary spheroids with small size will be observed 4-5 days after initial seeding. (IV) The spheres can be expanded for 10–14 days with mature colonies of large, phase bright, and spherical clusters. (b) Transferred method to culture hSKPs: (I) skin samples are cut into pieces, and the epidermis is removed after enzymatic digestion. (II) The pieces are mechanically dissociated into single cells. (III) Single cells are seeded into fibroblast medium with 10% FBS, and mature fibroblast colonies with spindle shape will be observed after 3–5 days. (IV) Cells from fibroblast culture are transferred into SKPs culture as the traditional procedure. After 4–14 days, mature spheres will be observed. SKPs: skin-derived precursors; hSKPs: human skin-derived precursors; bFGF: basic fibroblast growth factor; EGF: epidermal growth factor; FBS: fetal bovine serum.

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