How do you use culture mouse keratinocytes?

Culture the mouse KCs in KC growth medium (in humidified incubator with 5% CO2 at 37 °C) with daily medium change until the cells reach confluency. Immediately before the UVB irradiation, remove the growth medium and replace with 500 µL of PBS. Then subject the cells to 25 mJ/cm2 UVB or mock control (no UVB).

What type of cells are keratinocytes?

Keratinocytes are the primary type of cell found in the epidermis, the outermost layer of the skin. In humans, they constitute 90% of epidermal skin cells. Basal cells in the basal layer (stratum basale) of the skin are sometimes referred to as basal keratinocytes.

How do you isolate keratinocytes?

However, basing on the available publications, the best method for isolation of human keratinocytes is two-step overnight digestion in 1.45-2.5 U/mL dispase followed by short (up to 3 minutes) cleavage in 0.25% trypsin with subsequent pipetting of the epidermis.

What is keratinocyte apoptosis?

Keratinocyte (KC) apoptosis plays a critical role in regulating epidermal development and restraining carcinogenesis. Apoptosis balances proliferation to maintain epidermal thickness, contributes to stratum corneum formation and may eliminate pre-malignant cells.

Are basal cells parent cells?

In the airway epithelium, basal cells function as stem/progenitor cells that can both self-renew and produce differentiated secretory cells and ciliated cells.

How do you freeze primary keratinocytes?

Cryopreservation of Primary Keratinocytes Cryopreserve cells at a density of 0.5–2 × 106 cells/mL using a controlled-rate freezer or other appropriate device, then transfer to liquid nitrogen storage (vapor phase).

What is the function of a keratinocyte?

As the most dominant cell type constituting the epidermis, keratinocytes play multiple roles essential for skin repair. They are the executors of the re-epithelialization process, whereby keratinocytes migrate, proliferate, and differentiate to restore the epidermal barrier.

What is the origin of Merkel cells?

The origin of Merkel cells is unclear, as they share both epidermal and neuroendocrine features. Research suggests that they may be derived from pluripotential stem cells of the dermis or, as an alternative, from neural crest cells. Cytologic and immunohistochemical data support both contentions.

What is the normal function of basal cells?

Basal cells are found at the bottom of the epidermis — the outermost layer of skin. Basal cells produce new skin cells. As new skin cells are produced, they push older cells toward the skin’s surface, where the old cells die and are sloughed off.