Methods of culturing and modulating the growth of hair follicular stem cells

• US 5,556,783 A
• Filed: 07/01/1993
• Issued: 09/17/1996
• Est. Priority Date: 03/27/1991
• Status: Expired due to Term

• Current Assignee: New York University, Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: New York University, University of California Los Angeles (University of California)
• Inventors: Yang, Jing-Shan, Sun, Tung-Tien, Lavker, Robert M.
• Primary Examiner(s): Knode, Marian C.
• Assistant Examiner(s): DADIO, SUSAN
• Application Number:
  US08/086,199
• Time in Patent Office: 1,174 Days
• Field of Search: 435/240.2, 435/240.21, 435/240.23, 435/240.243, 435/240.25
• US Class Current: 435/366
• CPC Class Codes: C12N 2500/40 Nucleotides, nucleosides, bases cyclic nucleotides C12N2501/01, anti-neoplasic drugs C12N2501/06
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/70 Enzymes
• CPC Class Codes: C12N 2501/998 Proteins not provided for elsewhere
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 5/0628 Hair stem cells; Hair progenitors mesenchymal stem cells from hair follicles C12N5/0666
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics

Abstract

A method of culturing hair follicular stem cells by isolating a subpopulation of follicular keratinocytes from the upper portion of a hair follicle, dispersing the isolated keratinocytes into a single cell suspension, and growing the dispersed isolated keratinocytes in the presence of 3T3 feeder cells is provided. A method of evaluating the efficacy of agents for modulating the activity of bulge cell populations by exposing bulge cells to an agent to be tested and comparing the response of the test cells to established controls for bulge cells is also provided. Methods of modulating the activity of hair follicular stem cells by identifying a subpopulation of follicular keratinocytes from the upper portion of a hair follicle and contracting the cells with a selected agent to stimulate growth or to selectively kill the cells is also provided.

Claims

1. A method of culturing hair follicular stem cells comprising:

isolating from the upper portion of a hair follicle a subpopulation of follicular keratinocytes comprising hair follicular stem cells;

dispersing said isolated keratinocytes into a single cell suspension; and

growing said dispersed isolated keratinocytes together with 3T3 feeder cells or in a serum-free medium, thereby producing a hair follicular stem cell culture.

2. The method of claim 1 wherein said subpopulation of follicular keratinocytes is isolated by microdissection.

3. The method of claim 1, wherein said subpopulation of follicular keratinocytes comprises outer root sheath cells located in the midportion of the upper follicle at the arrector pili attachment site.

4. The method of claim 1 wherein said 3T3 feeder cells are mitomycin-treated.

5. The method of claim 4 wherein the cells are allowed to grow to about 70% confluence.

6. The method of claim 1 wherein the isolated keratinocytes are dispersed by incubating at about 37°

C. for about 30 minutes in about 0.25% trypsin.

7. The hair follicular stem cell culture produced by the method of claim 1.