Tissue repair using allogeneic dermal fibroblasts

• US 9,271,923 B2
• Filed: 03/08/2013
• Issued: 03/01/2016
• Est. Priority Date: 05/26/2005
• Status: Expired due to Fees

• Current Assignee: Intercytex Limited (Regenerative Medicine Assets Ltd.)
• Original Assignee: Intercytex Limited (Regenerative Medicine Assets Ltd.)
• Inventors: Leek, Mike, Kemp, Paul
• Primary Examiner(s): DAVIS, RUTH A
• Application Number:
  US13/791,177
• Publication Number:
  US 20130224158A1
• Time in Patent Office: 1,089 Days
• Field of Search: 435/325, 435/371
• US Class Current: 1/1
• CPC Class Codes: A61K 2800/91 Injection
• CPC Class Codes: A61K 8/981 of mammals or bird
• CPC Class Codes: A61K 8/985 Skin or skin outgrowth, e.g. hair, nails
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3839 characterised by the site of application in the body materials for artificial blood vessels A61L27/507; materials for use in artificial skin A61L27/60
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/00 Drugs for skeletal disorders
• CPC Class Codes: A61Q 19/08 Anti-ageing preparations

Abstract

The present invention provides a method for the repair of subcutaneous or dermal tissue in a subject, comprising in one aspect the injection of a suspension of allogeneic dermal fibroblasts into the subject. By injecting a suspension of allogeneic cells, the invention provides, for example, long-term augmentation of the subadjacent tissue without the disadvantages that accompany the preparation and/or use of presently available materials such as autologous fibroblasts cells or collagen.

According to a first aspect of the invention there is provided a method for the augmentation of subcutaneous or dermal tissue in a subject, which method comprises the steps of:

• (i) providing a suspension of allogeneic dermal fibroblasts; and
• (ii) injecting an effective volume of the suspension into tissue subadjacent to the subcutaneous or dermal tissue so that the tissue is augmented.

The method is preferably cosmetic.

Claims

1. A method for the augmentation of subcutaneous or dermal tissue in a subject, which method comprises the steps of:

(i) providing a suspension comprising allogeneic dermal fibroblasts in a preservation medium having a pH of about 7.6, an osmolality of about 360 and comprising the following components;

6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid, Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl⁻

, H₂PO₄⁻

, HCO₃⁻

, HEPES, Lactobionate, Sucrose, Mannitol, Glucose, Dextran-40, Adenosine and Glutathione; and

(ii) injecting an effective volume of the suspension into tissue subadjacent to the subcutaneous or dermal tissue so that the tissue is augmented.

2. The method of claim 1, wherein the method is cosmetic.

3. The method of claim 1, further comprising a step of identifying a defect that is susceptible to amelioration by augmentation of the subadjacent subcutaneous or dermal tissue.

4. The method of claim 3, wherein the defect is a rhytid, stretch mark, a depressed scar, a cutaneous depression of non-traumatic origin or an under-development of the lip.

5. The method of claim 1, in which the fibroblasts are passaged.

6. The method of claim 1, wherein the suspension further comprises fibrin.

7. The method of claim 6, wherein the fibrin is at a final concentration of 0.1 to 20 mg/ml.

8. The method of claim 6, wherein the fibrin is human fibrin.

9. The method of claim 1, wherein the suspension further comprises collagen and/or hyaluronic acid.

10. The method of claim 1, which further comprises the steps of:

a) obtaining a dermal biopsy from a source allogeneic to the subject;

b) passaging the dermal fibroblasts from the dermal biopsy in a culture medium comprising between 0.5% and 20% non-human serum, so as to provide dermal fibroblasts substantially free of adipocytes, keratinocytes and extracellular matrix; and

c) forming a suspension of the incubated fibroblasts in the preservation medium.

11. The method of claim 10, in which the dermal biopsy is from human neonatal foreskin.

12. The method of claim 10, in which the suspension is formed by scraping the fibroblasts and/or exposing the fibroblasts to a proteolytic enzyme and/or an animal product-free nonenzymatic alternative thereto.

13. The method of claim 1, in which the subject is human.

14. The method of claim 1, wherein between 10⁴ and 10⁸ fibroblasts are injected.

15. The method of claim 1, wherein a volume of about 1 ml of suspension is injected.

16. The method according to claim 1, wherein the fibroblasts are suspended in the preservation medium at a temperature of about 2°

C. to about 8°

C. for at least one hour prior to injecting.

17. The method of claim 1, wherein between 10⁶ and 10⁸ fibroblasts are injected.

18. The method of claim 1, wherein 1-5×

10⁷ fibroblasts are injected.

19. The method of claim 1, wherein 1-4×

10⁷ fibroblasts are injected.