Method and culture medium for ex vivo enrichment, expansion and long term culture of hair follicle stem cells

Prozesse und Methoden (inkl. Screening) : Lebens-Wissenschaften (inkl. Screening)
Nukleinsäure-, Protein-, Zell-bezogene Technologien : Zellen (bezogen)

Ref.-Nr.: 1013-5012-IKF

A method for de novo generation and long-term culture of epidermis-derived stems cells in feeder-free conditions and a special cell culture medium for in vitro tissue production, drug discovery and toxicity screenings, and medical applications.


Adult somatic stem cells (SCs) fuel tissue renewal, repair, and remodeling in mature organs. By tuning their proliferation rate to match the changing needs of their resident tissues, SCs maintain organ form and function.

Mammalian skin is characterized by a hair coat that maintains body temperature, homeostasis and serves a protective function. During postnatal life, mammalian skin and its hair coat are constantly renewed. SCs residing in the epidermis (referred to as epidermis-derived stem cells) such as interfollicular epidermal SCs or hair follicle stem cells (HFSCs) ensure the maintenance of adult skin homeostasis and trigger hair regeneration.

Recently, cell culture systems that support survival and expansion of SCs from mouse and human epithelial tissues other than the epidermis have been reported. In particular, no culture system for epidermis-derived stem cells is known that allows specific and targeted enrichment and expansion of these cells in vitro.

Therefore there is an urgent need to provide means and methods for ex vivo culturing and expansion of epidermis-derived stem cell.


Scientists of the Max Planck Institute for Biology of Ageing in Cologne developed a new method enabling the de novo generation, maintenance and enrichment of functionally competent epidermis-derived stem cells that display CD34+α6+ HFSCs characteristics. To achieve that they combined a 3-dimensional extracellular matrix (e.g. Matrigel) with different growth factors as well as a ROCK and a SSH inhibitor, and created an environment both necessary and sufficient to enable de novo generation, growth and expansion of cells with HFSCs features by mimicking a SC niche. Furthermore, using full-thickness skin reconstitution assay, they could show that cells cultured under above-described conditions retain their self-renewal capacity and multipotency leading to hair growth upon transplantation.

Based on this, a use of this cultured HFSCs in tissue transplantation and/or in the treatment of dermal burns, or treatment of conditions where areas of skin have been removed due to surgical operation, biopsy, burn or trauma as well as under conditions where the regenerative capacity of the skin is compromised such as chronic wound or baldness is possible. Furthermore, cultured HFSCs can be used as a screening platform to identify compounds that enhance the function or de novo generation of HFSCs to be used in regenerative medicine or cosmetic applications.

Fig.1: Establishment of a HFSC culture system and demonstration of self-renewing capacity and multipotency.

A) Epidermal cells grown for 14days in the indicated conditions. 3C: 3D-Matrigel in basal medium with growth factors and inhibitors. 2D: 2D in 3C medium. Scale bars 30µm.
B) Immunofluorescence analysis of 3C cultures showing the expression of the HFSC markers CD34 and SOX9 in a subset of cells. Scale bars 25µm.
C) Colony-forming assay shows increased proliferative potential of cells pre-cultured in 3C compared to freshly isolated epidermal cells (Epi d0; control; mean +/- SEM; n=5; *P<0.05, Mann-Whitney U-test). 3C: 3D-Matrigel in basal medium with growth factors and inhibitors.
D) Full-thickness skin reconstitution assay with freshly isolated epidermal cells (Epi d0) or 3C cultures shows that cells cultured in 3C retain their self-renewal capacity and multipotency. A representative recipient of four mice/conditions is shown. Right panel shows quantifications of hair follicle density. Note that 3C cultures give rise to more hair compared to controls (mean +/- SEM; n=4; *P<5.05; Mann-Whitney U-test).

In addition to this, this technology includes the development of culture conditions of expansion and maintenance of epidermis-derived cancer stem cells (CSCs), allowing screens for compounds that affect the function of these cells potentially to be used in cancer therapy.


The present technology relates to a method for ex vivo de novo generation and/or expansion of epidermis-derived stem cells in a 3D-extracellular matrix and a special cell culture medium for in vitro tissue production, in vitro drug discovery and toxicity screenings and medical applications.

We are now looking for a collaboration- and/or licensing partner who is interested in this approach and eager to develop these scientific achievements into a marketable good.

Patent Information

  • PCT application filed: 04.10.2016
  • PCT-Patent Application PCT/EP2016/073675


Chacón-Martínez C.A. et al. (2017): Hair follicle stem cell cultures reveal self-organizing plasticity of stem cells and their progeny The EMBO Journal. 36: 151-164. Selected as F1000Prime publication.

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Dr. Ingrid Kapser-Fischer

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