iPSC-derived skin cells
Using three unique iPSC lines from donors of different ethnicities we have developed a wide range of skin cells including iPSC-derived Melanocytes, iPSC-derived Sebocytes, and iPSC-derived Keratinocytes. Each of these cells are consistent and reproducible as they display high purity, low batch to batch variation and are available in large batch sizes, making them ideal for toxicology investigations and drug discovery.
Our iPSC-derived Melanocytes express the characteristic melanocyte-specific markers, PMEL17, TYRP1 and MITF. Human iPSC-derived Melanocytes exhibit melanin synthesis and the ability to transfer melanosomes to keratinocytes making them excellent models for in vitro studies into pigmentation and the development of photoprotective drugs. We also offer a fully optimized melanocyte culture system that includes Melanocyte Proliferation Medium which can be used for expansion and a Melanocyte Growth Medium which ideal for assays.
Our Human iPSC-derived Sebocytes express the major sebocyte markers MUC1 and KRT7 along with the expected accumulation of lipids. These cells are physiologically similar to human sebocytes as they express key functional genes involved in lipid synthesis and differentiation and they exhibit the expected response to treatment of linoleic acid and testosterone. Human iPSC-derived Sebocytes lend themselves to research into skin disorders such as acne and dermatitis. Our Human iPSC-derived Sebocytes are supplied with supplements that support the generation of sebocytes with a low or high lipid content depending on assay requirements.
Our iPSC-derived Keratinocytes are still in development and are due for release in 2018!
Human iPSC-derived melanocytes
Human iPSC-derived sebocytes
|Cat. No.||Product Name||Quantity|
|ax0530||Human iPSC-Derived Sebocytes (Caucasian)||2 million and 2x supplements|
|ax0531||Human iPSC-Derived Sebocytes (African)||2 million and 2x supplements Pre-order now - Coming Soon!|
|ax0532||Human iPSC-Derived Sebocytes (Asian)||2 million and 2x supplements Pre-order now - Coming Soon!|
Human iPSC-derived keratinocytes
Phase contrast melanocytes
Phase contrast images show the difference between the three available phototypes III-IV, IV-V and VI.
Phase contrast images of Axol Human iPSC-Derived Melanocytes. Human iPSC-derived melanocytes from three different phototypes (African (VI), Asian (IV-V) and Caucasian (III-IV)) are cultured in Melanocyte Proliferation Medium at a density of 20,000 cells/cm2. The iPSC-derived melanocytes form an adherent, confluent monolayer and display a star-like morphology with a gradual increase in pigmentation over number of days in culture.
Axol iPSC-Derived Melanocytes express key functional melanogenic proteins that are involved in the regulation and production of the pigment melanin.
Immunocytochemistry analysis of iPSC-Derived Melanocytes. Human iPSC-Derived Melanocytes show characteristic marker expression in the correct subcellular localization when stained for PMEL17, TYRP1 and MITF (green) and counterstained with DAPI (blue).
Flow cytometry melanocytes
Axol Human iPSC-derived Melanocytes show high purity when analysed for the premelansome marker, PMEL17. Axol Human iPSC-Derived Melanocytes were thawed and cultured for one passage until 90% confluent before flow cytometry analysis was performed. iPSC-derived melanocytes from the three ethnicities show over 95% purity. The graph shows isotype control in black and PMEL17 antibody in blue.
Transfer and uptake of melanosomes
Axol iPSC-derived Melanocytes in co-culture with primary keratinocytes show the transfer of melanin in melanosomes to keratinocytes.
Functional analysis of human iPSC-Derived Melanocytes. Human iPSC-derived Melanocytes from all three ethnicities show characteristic perinuclear organization of melanosomes (A-C). Human iPSC-derived Melanocytes were amplified in Melanocyte Proliferation Medium for 1 passage before placing into co-culture with human primary keratinocytes (passage 3) and changing to co-culture medium. The co-culture of iPSC-derived melanocytes with keratinocytes resulted in a maturation of melanoctye morphology from a star-like morphology to an extended spindle morphology with dendrites extending towards the keratinocytes to enable melanosome transfer.
The circles dictate regions where the transfer of TYRP1-labelled melanosomes (red) to primary keratinocytes stained with Cytokeratin-14 (green) can be observed (A-C). Intracellular fluorescent dye carboxyfluorescein succinimidyl ester (CFSE, green) was used to stain the cytoplasm of primary keratinocyte in order to monitor the uptake of TYRP1-labelled melanosomes (red) from iPSC-Derived Melanocytes (Caucasian) (D). All counterstained with Hoechst (blue).
Melanin synthesis melanocytes
Axol Human iPSC-derived Melanocytes produce melanin when stimulated by α-MSH or UV. Human iPSC-derived Melanocytes treated with α-MSH (5 µM) for 48 hours showed a 75% significant increase in melanin content (n=3 p<0.05 *) (A). iPSC-derived Melanocytes were exposed to an increasing irradiation dose of UV (0, 20, 60 mJ/cm2). Melanin pigmentation/ million cells was measured 3 days after UV treatment, melanin content increased by 4-fold and enhanced pigmentation was observed by phase contrast (B-C).
Phase contrast sebocytes
Axol Human iPSC-derived Sebocytes have expected epithelial morphology.
Phase contrast images of Axol Human iPSC-Derived Sebocytes. Human iPSC-derived Sebocytes (Caucasian) were seeded at a density of 25,000 cells/cm2 and cultured for 5 days. Over 5 days lipid droplet accumulation becomes apparent (yellow arrows) within the iPSC-Derived sebocytes which results in a heterogeneous epithelial morphology.
Axol iPSC-derived Sebocytes express key sebocyte markers MUC1 and KRT7.
Immunocytochemistry analysis of hiPSC-Derived Sebocytes. Human iPSC-derived Sebocytes show characteristic marker expression the correct subcellular localization when stained for differentiation specific markers, MUC1 (green) and KRT7 (red), counterstained with Hoechst (blue). The flourescent probe, BODIPY, shows lipid accumulation in the majority of iPSC-derived sebocytes.
Flow cytometry sebocytes
Axol Human iPSC-Derived Sebocytes show high purity when analysed for the sebocyte marker, KRT7 (Keratin 7). Flow cytometry analysis of human iPSC-derived sebocytes was performed after 5 days in culture (3 days in Sebocyte Basal Medium with Amplification Supplement and 2 days in Sebocyte Basal Medium with Muturation Supplement). Expression of KRT7 is greater than 90% purity in Axol Human iPSC-derived Sebocytes.
Gene expression sebocytes
Development of functional markers in Axol Human iPSC-Derived Sebocytes. Human iPSC-derived sebocytes were analysed for their gene expression days 3 (d3) and 5 (d5) in culture in comparison to human primary keratinocytes (passage 3) using quantitative reverse transcription PCR (RT-qPCR). After 5 days in culture key functional genes involved in lipogenesis (PPAR𝜸 and FADS2, sebocyte regulation (androgen receptor, AR) and sebocyte differentiation (MC-5R) were all strongly expressed.
Lipid synthesis sebocytes
Linoleic acid and testosterone are known to stimulate lipid production.
Response of Axol Human iPSC-derived Sebocytes to linoleic acid treatment. Human iPSC-derived sebocytes treated with increasing concentrations of linoleic acid (LA) for 24 hours showed a significant does-dependent increase in lipid accumulation, measured by the fluorescent probe BODIPY. n=3
Response of Axol Human iPSC-derived Sebocytes to testosterone treatment. The treatment of Human iPSC-derived sebocytes with testosterone (10 µM) for 96 hours significantly induced a 2-fold increase in lipid content. Further treatment with testosterone and the 5α- reductase inhibitor, Finasteride (10 µM), for 96 hours resulted in a significant reduction in lipid accumulation (BODIPY staining). n=3