Vitro cellular interaction with amnion membrane substrate

• US 4,446,234 A
• Filed: 10/23/1981
• Issued: 05/01/1984
• Est. Priority Date: 10/23/1981
• Status: Expired due to Term

• Current Assignee: United States Department Of Health And Human Services
• Original Assignee: United States Department Of Health And Human Services
• Inventors: Liotta, Lance A., Russo, Raimondo
• Primary Examiner(s): Warden, Robert J.
• Application Number:
  US06/314,477
• Time in Patent Office: 921 Days
• Field of Search: 435/4, 435/6, 435/29, 435/32, 435/33, 435/35, 435/240, 435/241, 435/284, 435/285, 435/286
• US Class Current: 435/29
• CPC Class Codes: A61L 2430/40 Preparation and treatment of biological tissue for implantation, e.g. decellularisation, cross-linking
• CPC Class Codes: A61L 27/3604 characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: C12N 2533/92 Amnion; Decellularised dermis or mucosa
• CPC Class Codes: C12N 5/0068 General culture methods using substrates for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/067 Hepatocytes
• CPC Class Codes: G01N 33/5011 for testing antineoplastic activity
• CPC Class Codes: G01N 33/5091 for testing the pathological state of an organism

Abstract

Amnion membranes derived from mammalian placentas are employed as the substrate tissue in in vitro assay methods for evaluating cellular interaction with natural barrier tissues. The amnion is also employed as a growth substrate, particularly for the culture of cells difficult to culture on conventional substrates. A diagnostic apparatus including an amnion membrane is also disclosed.

Claims

1. In an in vitro assay method for evaluating cellular interaction with natural barrier tissue of the type wherein living cells are interacted in vitro with a substrate tissue, the improvement comprising employing a mammalian amnion membrane as the substrate tissue.

2. The invention of claim 1, wherein the cellular interaction to be evaluated comprises cellular invasion or penetration of the natural barrier tissue.

3. The invention of claim 1, wherein the amnion is derived from a human placenta.

4. The invention of claim 3, wherein the cells are inflammatory cells and are interacted in the presence of a substance from a biological source.

5. The invention of claim 4, wherein the cells are polymorphonuclear leucocytes.

6. The invention of claim 1, wherein the living cells are malignant tumor cells.

7. The invention of claim 6, wherein the tumor cells are interacted in the presence of a pharmaceutical composition.

8. The invention of claim 1, wherein the cells are normal migratory cells.

9. The invention of claims 6 or 8, wherein the cells are interacted in the presence of a chemoattractant.

10. The invention of claim 1 wherein the amnion connective tissue components are radioactively labeled.

16. The invention of claims 1, 2, 3, 6, 8, 7, 4, 14, or 15, wherein the amnion membrane is wholly or partially denuded of its associated epithelium.

11. A method for culturing cells in vitro comprising interacting living cells with a mammalian amnion membrane growth substrate.

12. The invention of claim 11, wherein the cells are interacted with the growth substrate in the presence of a serum-free culture medium.

13. The invention of claim 12, wherein the cells are hepatocytes.

14. A diagnostic apparatus comprising a chamber including a mammalian amnion membrane dividing the chamber into upper and lower compartments, and support means for supporting the amnion membrane within the chamber.

15. The invention of claim 14, wherein the support means comprises an upper ring, a lower ring, and securing means for securing the rings to form said chamber, and wherein the amnion is clamped between said rings to divide the chamber into upper and lower compartments.