System and method for transplantation of cells

• US 5,423,778 A
• Filed: 06/11/1992
• Issued: 06/13/1995
• Est. Priority Date: 12/14/1989
• Status: Expired due to Term

• Current Assignee: Applied Tissue Technologies LLC
• Original Assignee: Elof Eriksson
• Inventors: Vogt, Peter M., Eriksson, Elof
• Primary Examiner(s): Rosenbaum, C. Fred
• Assistant Examiner(s): Alexander, V.
• Application Number:
  US07/897,357
• Time in Patent Office: 1,097 Days
• Field of Search: 604/304-308, 604/890.1, 604/891.1, 623/15, 424/DIG. 13, 424/574, 424/422, 424/423, 128/888, 435/41, 435/172.1, 435/240.1, 435/240.2, 435/240.21, 435/240.23, 435/240.31, 435/240.241, 435/240.243, 435/287, 435/289
• US Class Current: 604/305
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 35/36 Skin; Hair; Nails; Sebaceous glands; Cerumen; Epidermis; Epithelial cells; Keratinocytes; Langerhans cells; Ectodermal cells islets of Langerhans A61K35/39
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 9/0021 Intradermal administration, e.g. through microneedle arrays, needleless injectors mechanical aspects A61M
• CPC Class Codes: A61K 9/1676 having a drug-free core with discrete complete coating layer containing drug adsorbates of liquid drug formulations on inert powders without simultaneous granulation step A61K9/141; with further drug-free outer coating A61K9/5078; drug conjugated to non-active particles A61K47/6921
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/60 Materials for use in artificial skin
• CPC Class Codes: A61M 2037/003 having a lumen
• CPC Class Codes: A61M 2037/0038 having a channel at the side surface
• CPC Class Codes: A61M 2037/0061 Methods for using microneedles
• CPC Class Codes: A61M 37/00 Other apparatus for introducing media into the body for reproduction or fertilisation A61B17/425; apparatus for iontophoresis or cataphoresis A61N1/30; Percutany, i.e. introducing medicines into the body by diffusion through the skin salt baths A61H33/04
• CPC Class Codes: A61M 37/0015 by using microneedles
• CPC Class Codes: A61M 37/0076 Tattooing apparatus apparatus for marking animals A01K11/00; vaccine applicators having needles or other puncturing means A61B17/205
• CPC Class Codes: A61M 39/0247 Semi-permanent or permanent transcutaneous or percutaneous access sites to the inside of the body peritoneal dialysis catheters A61M1/285; tracheostomy devices A61M16/0465; measuring pressure within the body A61B5/03; colostomy devices A61F5/445; gastrotomy feeding tubes A61J15/0015; means for fixing a feeding tube outside of the body A61J15/0053
• CPC Class Codes: A61M 5/3287 Accessories for bringing the needle into the body; Automatic needle insertion A61M5/20, A61M5/31525 take precedence
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2799/027 where the vector is derived from a retrovirus
• CPC Class Codes: C12N 5/0062 General methods for three-dimensional culture
• CPC Class Codes: C12N 5/0629 Keratinocytes; Whole skin

Abstract

Gene transfer of genetic material with viral vectors or plasmid, in combination with a wound treatment chamber, into keratinocytes, especially those including a high percentage of epidermal stem cells, has been demonstrated to be particularly effective as a means of implanting genetically engineered cells and obtaining long term survival. By employing the wound chamber system, direct in vivo gene transfer can also be done to exposed cells in an open wound. Skin stem cells which are located in the hair follicles are used to greatly enhance long term survival. The use of the wound chamber system for gene transfer also allows non-invasive assessment of the success of transfer by assaying for the presence of the expressed protein in wound fluid, in contrast to the prior art use of invasive techniques, such as biopsies, in order to achieve the same assessment of early expression. A wide variety of proteins and materials can be expressed, either for secretion into the general blood and lymphatic system, or to alter the properties of the protein, for example, to not express proteins eliciting an immune response against the transplanted cell.

Claims

1. A method for culturing cells at a site on and within a patient comprising attaching to the surface of the patient and at the site on a patient where cells are to be grown a chamber formed of a flexible, moisture and gas impermeable material containing a fluid medium suitable for culturing of cells and open to the patient so that the medium is in contact with the patient.

2. The method of claim 1 further comprising adding cells to be cultured into the chamber.

3. The method of claim 2 wherein the cells are selected from the group consisting of keratinocytes and epidermal stem cells.

4. The method of claim 3 wherein the epidermal stem cells are isolated by removing the cells from hair follicles on the underside of partial thickness skin flaps.

5. The method of claim 2 wherein the cells are engineered to express exogenous genetic material.

6. The method of claim 5 wherein the cells are engineered to express proteins selected from the group consisting of hormones, immunomodulators, clotting factors, antigens, and antibodies.

7. The method of claim 2 wherein the cells are engineered to not express proteins eliciting an immune reaction against the cells.

8. The method of claim 7 wherein the cells do not express HLA antigens.

9. The method of claim 2 further comprising introducing exogenous genetic material into the cells in the chamber.

10. The method of claim 1 further comprising controlling culture variables in the chamber.

11. The method of claim 10 wherein the variables are selected from the group consisting of pH, gas concentrations, ion concentrations, and temperature.

12. The method of claim 1 further comprising adding compounds to the chamber selected from the group consisting of antibiotics, anesthetics, chemotherapeutic agents, immunomodulators, and growth factors.

13. A method for increasing the yield of genetically engineered cells in a patient comprising the steps of selecting in vivo epidermal stem cells as the cells to be engineered and introducing genetic material into said epidermal stem cells.

14. The method of claim 13 further comprising selecting the cells from the same species as the patient.

15. A method according to claim 13 comprising the further steps of covering the genetically engineered stem cells with a chamber having an opening securable at the periphery to the skin of the patient and being formed of a flexible, moisture and gas impermeable material and sealing the periphery of the chamber to the skin of the patient.

16. A method for genetically engineering cells in a patient, comprising

attaching to the surface of the patient and at a site where cells are to be engineered a chamber formed of a flexible, moisture and gas impermeable material, the chamber containing a fluid medium suitable for culturing cells and open to the cells to be engineered, and

introducing genetic material into the cells covered by the chamber.

17. The method of claim 16 wherein the cells are keratinocytes.

18. The method of claim 16 wherein the genetic material is introduced into the cells using a vector selected from the group consisting of viral vectors and plasmids.

19. The method of claim 16 wherein genetic material is introduced into the cells, further comprising controlling temperature, pH, growth of microorganisms, gas and ion concentrations.

20. The method of claim 16 further comprising providing in the chamber compounds selected from the group consisting of antibiotics, anesthetics, chemotherapeutic agents, immunomodulators, and growth factors.

21. A method for increasing the yield of genetically engineered cells implanted in a patient, comprising the steps of selecting epidermal stem cells as the cells to be engineered, covering the implanted stem cells with a chamber, and introducing genetic material to the cells within said chamber, said chamber having an opening securable at the periphery to the skin of the patient and being formed of a flexible, moisture and gas impermeable material wherein said chamber contains the genetic material and the cells to be genetically engineered.

22. A method according to claim 21 wherein the step of selecting epidermal stem cells includes the step of exposing the basal layer of the patient'"'"'s epidermis for the stem cells.