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Exploring the Power of Human Epidermal Keratinocytes in Research and Medicine

Human Epidermal Keratinocytes (HEK) are ideal cell platforms to study wound healing, toxicology, skin biology/diseases, human papillomaviruses (HPV) infection, regenerative medicine and dermatological/cosmetic research.  HEKs are also used as in vitro human skin models to evaluate anti-inflammatory effects of cosmetic formulations. Epidermis, the outermost skin layer, is composed of 90% HEK. It acts as a physical barrier, the intermediate filament keratin. Keratinocytes receive melanin from melanocytes and store it for protection from UV radiation. HEK cells are either primary cells isolated from tissue biopsies or can be derived from induced pluripotent stem cells (iPSCs).

Primary Human Epidermal Keratinocytes (HEK)

Human Epidermal Keratinocytes
FIGURE 1: Isolation of primary Human Epidermal Keratinocytes from skin tissue (Image Source: PMID: 35199036)

HEKs are generally isolated from human skin tissues from surgical waste materials with donor consent. HEKs are isolated by enzymatic treatment with dispase or trypsin after separating epidermis from dermis. It can also be isolated by explant culture methods. Isolated keratinocytes are cultured in specific media with growth factors e.g. keratinocyte growth factors (KGF), Epidermal Growth Factor (EGF) etc. This specialized media supports proliferation and maintenance. HEK also required a feeder layer or surface coating for enhanced cell attachment and growth.

iPSC-Derived Keratinocytes

Induced pluripotent stem cells (iPSCs) are generated from human somatic cells e.g. Peripheral Blood Mononuclear Cells (PBMCs), fibroblast etc. using Yamanaka factors OCT4, SOX2, KLF4, C-MYC. iPSCs are sequentially differentiated towards keratinocytes using growth factors and signaling molecules through ectodermal lineage specification by mimicking embryonic epidermal development.

 

Characterization of HEK

iPSC-Derived Keratinocytes
FIGURE 2: Directed differentiation of human iPSCs towards keratinocytes (Image Source: 31058887)
  • Morphological Analysis: HEK exhibits a cobblestone-like morphology. 
  • Molecular characterization: Expression of specific keratinocyte markers such as Keratin 5, Keratin 14, Keratin 10, and Keratin 1 is assessed using immunocytochemistry and flow cytometry. Involucrin, Loricrin, and filaggrin also confirm the differentiated phenotype of HEK. α6β4 integrin and E-Cadherin also play a crucial role in maintaining the integrity of the epidermal layer.
  • Functional Characterization: Barrier function assay is used to assess the integrity and permeability of the keratinocyte layer.

Applications of HEK

  • Disease Modeling: HEK and iPSC-derived keratinocytes are used to study disease mechanisms and their therapeutic interventions by various skin disease modeling, e.g. psoriasis, atopic dermatitis, eczema, epidermolysis bullosa etc.
  • Drug Screening and Toxicology: Keratinocytes serve as a model for efficacy, toxicology and safety assessment of dermatological drugs, cosmetics and other topical ointments. 
  • Regenerative Medicine: Keratinocytes are used to generate skin grafts to treat chronic wounds like burns and other skin injuries.
  • Gene Editing and Therapy: CRISPR/Cas9 editing tools are used to treat various genetic skin disorders, e.g. Ichthyosis, vitiligo etc.
  • Personalized Therapy: iPSCs-derived keratinocytes enable facilitating personalized therapeutic strategies, improving treatment outcomes.

Conclusion

HEK are invaluable tools in dermatological research and regenerative medicine. Their ability to model diseases, screen drugs, and contribute to personalized medicine underscores their significance. At Kosheeka, we are committed to advancing these technologies to support groundbreaking research and therapeutic developments. HEKs are amenable to high throughput drug screening for toxicology and safety studies. For more information about our customized cell culture services and to learn how we can support your research needs, please contact us.

Frequently Asked Questions (FAQ):

Q.1 What are Human Epidermal Keratinocytes (HEKs)?
Human Epidermal Keratinocytes (HEKs) are the primary type of cells found in the epidermis, the outermost layer of the skin. They play a crucial role in forming a physical barrier against environmental damage, pathogens, and UV radiation.
Q.2 Why are HEKs Considered Ideal for Studying Various Biological Processes and Applications
HEKs are ideal for studying wound healing, toxicology, skin biology, diseases, HPV infection, regenerative medicine, and dermatological/cosmetic research because they accurately represent the human skin environment. Their ability to mimic the skin's response to various stimuli makes them excellent models for these studies.
Q.3 How are Primary Human Epidermal Keratinocytes Isolated?
Primary HEKs are isolated from human skin tissues obtained from surgical waste materials with donor consent. The isolation process involves enzymatic treatment with dispase or trypsin to separate the epidermis from the dermis. They can also be isolated using the explant culture method and cultured in specific media with growth factors to support proliferation and maintenance.
Q.4 What are iPSC-Derived Keratinocytes?
iPSC-Derived Keratinocytes are keratinocytes generated from induced pluripotent stem cells (iPSCs). iPSCs are created from human somatic cells, such as Peripheral Blood Mononuclear Cells (PBMCs) or fibroblasts, using reprogramming factors like OCT4, SOX2, KLF4, and C-MYC. These iPSCs are then differentiated into keratinocytes using growth factors and signaling molecules that mimic embryonic epidermal development.
Q.5 How are HEKs Characterized?
  • Morphological Analysis: HEKs exhibit a cobblestone-like morphology.
  • Molecular Characterization: Specific markers like Keratin 5, Keratin 14, Keratin 10, and Keratin 1 are assessed using immunocytochemistry and flow cytometry. Markers such as Involucrin, Loricrin, and Filaggrin confirm the differentiated phenotype. α6β4 integrin and E-Cadherin play crucial roles in maintaining epidermal integrity.
  • Functional Characterization: Barrier function assays assess the integrity and permeability of the keratinocyte layer.
Q.6 What are the Applications of HEKs in Research and Medicine?
  • Disease Modeling: HEKs and iPSC-derived keratinocytes are used to model various skin diseases and study therapeutic interventions for conditions like psoriasis, atopic dermatitis, eczema, and epidermolysis bullosa.
  • Drug Screening and Toxicology: Keratinocytes serve as models for testing the efficacy, toxicology, and safety of dermatological drugs, cosmetics, and topical ointments.
  • Regenerative Medicine: HEKs are used to generate skin grafts for treating chronic wounds, burns, and other skin injuries.
  • Gene Editing and Therapy: CRISPR/Cas9 tools are used to treat genetic skin disorders like ichthyosis and vitiligo.
  • Personalized Therapy: iPSC-derived keratinocytes facilitate personalized therapeutic strategies, improving treatment outcomes.
Q.7 Why are HEKs Invaluable in Dermatological Research and Regenerative Medicine?
HEKs are invaluable due to their ability to accurately model human skin responses, which is essential for studying disease mechanisms, screening drugs, and developing personalized medicine. Their extensive application potential makes them crucial for advancements in dermatological research and regenerative medicine.
Q.8 How does Kosheeka Support Research Involving HEKs?
Kosheeka is committed to advancing technologies related to HEKs to support groundbreaking research and therapeutic developments. They offer customized cell culture services and provide comprehensive support for research needs. For more information, interested parties can contact Kosheeka or visit their website.

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