Methods for producing hair microfollicles and de novo papillae and their use for in vitro tests and in vivo implantations

Patent metadata

Field	Value
Publication number	US-9764064-B2
Application number	US-201414468494-A
Country	US
Kind code	B2
Filing date	Aug 26, 2014
Priority date	Mar 28, 2008
Publication date	Sep 19, 2017
Grant date	Sep 19, 2017

Abstract

The present invention relates to a method for producing hair microfollicles comprising the steps of: a) providing de novo papillae, b) providing other cell populations selected from the group of fibroblasts, keratinocytes and melanocytes, and co-culturing the de novo papillae with at least one other cell population in non-adherent culture vessels. The present invention relates also to methods of producing de novo papillae usable in said method for producing hair microfollicles.

First claim

We claim:

1. A de novo papilla consisting of a cell aggregate of mammalian dermal hair papilla fibroblasts (DPF), the cell aggregate exhibiting the shape and the size of a physiological dermal papilla (DP) of a hair follicle after isolation and being coated with an extracellular matrix protein, wherein the de novo papilla is produced by a method comprising the steps of:
(a) providing at least one dermal papilla (DP) from at least one mammal hair follicle;
(b) isolating dermal hair papilla fibroblasts (DPFs) from the DP by mechanically fixing said DP at the surface of a cell culture vessel, whereby the basal lamina is perforated to allow said DPFs migrating out;
(c) expanding the isolated DPFs in monolayer culture without collagen coating, wherein said DPFs are passaged at least once;
(d) condensing the expanded DPFs in non-adhesive culture vessels in a cell concentration per vessel surface of 1,000 to 100,000 DPFs/cm 2 into cell aggregate and differentiating the cell aggregate into cell aggregates that exhibit the size and shape of the physiological DP in a non-adhesive culture vessel; and
(e) coating the cell aggregates produced in (d) with the extracellular matrix protein, thereby producing the de novo papilla.

2. The de novo papilla of claim 1 , wherein in step (d) of the method the expanded DPFs are condensed for at least 48 h.

3. The de novo papilla of claim 1 , wherein in step (d) of the method the expanded DPFs are condensed for 2 to 21 or 3 to 15 days.

4. The de novo papilla of claim 1 , wherein in step (d) of the method non-inductive DPFs are condensed.

5. The de novo papilla of claim 1 , wherein the extracellular matrix protein is collagen IV, fibronectin and/or laminin.

Assignees

• Univ Berlin Tech

Inventors

• Lindner Gerd
• Lauster Roland

Classifications

• A61P17/00

  Drugs for dermatological disorders · CPC title

• A61P17/14

  for baldness or alopecia · CPC title

• A61L27/3886

  comprising two or more cell types · CPC title

• C12N2533/52

  Fibronectin; Laminin · CPC title

• C12N2533/54

  Collagen; Gelatin · CPC title