Artificially grown, animal-free skin models are an important tool for simulating the effects of different conditions on epidermal tissues. In order to effectively culture such models, researchers must successfully induce keratinocytes to differentiate into the various layers of epidermis, requiring direct exposure to air as well as culture media that is used to support cell growth and differentiation.
Relevant applications and testing protocols:
Animal-free skin cell testing
Cosmetics product testing
Dermal toxicity assessment
Skin care product testing
Human skin cell growth
In vitro 3D epidermal tissue culture
Small-scale 3D skin growth
Human epidermal keratinocytes
Growing tissue with an air-liquid interface
Human epidermal reconstruction modeling
Discover a highly effective, time-saving method for culturing 3D skin epidermal tissue models
The Thermo Scientific Nunc Cell Culture Insert System offers a highly reliable, effective and efficient solution for culturing 3D epidermal skin models used in numerous applications—from skin care product testing to clinical screening of compounds and formulations.
The system for growing 3D skin tissue models includes:
A 24-well cell culture treated plate, preloaded with 4μm pore size polycarbonate membrane inserts
The proprietary 24-well carrier plate, designed to enable three different hanging height adjustments to accommodate required liquid-air interface culture and variable cell feeding protocols
How does skin tissue culture work?
The Nunc Cell Culture Insert System offers an effective method for culturing 3D skin epidermal tissue. The adjustable-height capabilities of the Thermo Scientific Nunc Carrier Plate simplify the experimental protocol, extending feeding intervals and saving time and labor in the process.
Initial experiments on cell attachment and expansion indicate that Nunc Cell Culture Inserts are an excellent growth substrate for human epidermal keratinocytes.
In the image below, the hematoxylin and eosin (H&E) stain shows the stratification of the epidermal skin model cultured using the Nunc Cell Culture Insert System.
MTT assay after two weeks of air-liquid interface culture of Human Epidermal Keratinocyte (HEKa) showed good viability in Nunc Cell Culture Inserts.
Assessment of the integrity using transepithelial electrical resistance (TEER) measurement after initial seeding of the HEKa cells in the 24-well inserts.
Depiction of cell culture inserts hanging in carrier plate, showing three different hanging positions
Dimensions given represent the height of the growth surface of the cell culture insert from the interior bottom of the well. Dimensions in figure are rounded to the tenth of a millimeter.
Tissue sections photographed at 400X magnification
We took advantage of the versatility of the carrier plate and tested different insert hanging positions, and corresponding increases in media volume, for different media change intervals of every 2, 3 or 4 days. This figure shows H&E stains of stratified skin model tissue after 12 days of air-liquid interface culture with media changes at varying intervals in carrier plates that suspended inserts above the wells at three different hanging heights. The higher hanging position with longer intervals worked as effectively as the lower hanging position with shorter intervals, saving time and labor during the skin tissue establishment.
A) Low hanging position: 0.5mL media per well, fed every 2 days.
B) Middle hanging position: 1.0mL media per well, fed every 3 days.
C) High hanging position: 1.5mL media per well, fed every 4 days.
What products do I need to buy for epidermal tissue culture?