Potentially regenerative cells and functional tissue-organs in vitro

• US 20060292692A1
• Filed: 08/30/2006
• Published: 12/28/2006
• Est. Priority Date: 09/27/2002
• Status: Abandoned Application

• Current Assignee: Rongxiang Xu
• Original Assignee: Rongxiang Xu
• Inventors: Xu, Rongxiang
• Application Number:
  US11/513,442
• Publication Number:
  US 20060292692A1
• Time in Patent Office: Days
• US Class Current: 435/325
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/76 from plants
• CPC Class Codes: C12N 2500/82 from invertebrates
• CPC Class Codes: C12N 2501/00 Active agents used in cell culture processes, e.g. differentation
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/999 Small molecules not provided for elsewhere
• CPC Class Codes: C12N 5/0018 Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0617 Thyroid and parathyroid glands
• CPC Class Codes: C12N 5/0619 Neurons
• CPC Class Codes: C12N 5/0627 Hair cells
• CPC Class Codes: C12N 5/0657 Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0669 Bone marrow stromal cells; Whole bone marrow isolated stem cells from bone marrow C12N5/0647, C12N5/0663
• CPC Class Codes: C12N 5/067 Hepatocytes
• CPC Class Codes: C12N 5/0676 Pancreatic cells
• CPC Class Codes: C12N 5/0679 Cells of the gastro-intestinal tract
• CPC Class Codes: C12N 5/0686 Kidney cells

Abstract

Compositions and methods are provided for culturing in vitro potentially regenerative cells (PRCs) from which functional tissue-organs are regenerated. In one aspect of the invention, a tissue culture medium is provided which comprises at least 50% of water and a sterol compound that is dissolved in a fatty acid-containing oil at a concentration at least 0.1% by weight based on the weight of the oil and added to the water. The culture medium can be used to culture PRCs that are isolated from the body of a mammal to generate functional tissue-organs in vitro with substantially the same physiological structure and function as the corresponding ones existing in vivo and in situ. The cultured PRCs, tissues, and tissue-organs can serve as valuable models for scientific investigation in life sciences, nutraceutical discovery, drug screening, pharmacokinetic studies, medical devices and tissue/organ transplantation.

Claims

1. Isolated potentially regenerative cells from a predetermined site of the body of a mammal, wherein the isolated potentially regenerative cells, when cultured in a culture medium under suitable conditions, are capable of being activated to continuously proliferate and differentiate to form a tissue-organ which shares substantially the same physiological structure and at least one physiological function with that of the corresponding tissue in situ and in vivo.

2. The isolated potentially regenerative cells of claim 1, wherein the culture medium is formed by the act comprising adding fatty acid-containing oil, in which a sterol compound of at least 0.1%, and wax of 1-20% by weight based on the total weight of the oil are dissolved or mixed, to a culture medium containing at least 50% of water.

3. The isolated potentially regenerative cells of claim 2, wherein the concentration of the oil in the tissue culture medium is about 1-50% by weight.

4. The isolated potentially regenerative cells of claim 2, wherein the concentration of sterol compound dissolved in the oil is about 0.5-40% based on the weight of the oil.

5. The isolated potentially regenerative cells of claim 2, wherein the concentration of sterol compound dissolved in the oil is about 1-20% based on the weight of the oil.

6. The isolated potentially regenerative cells of claim 2, wherein the concentration of sterol compound dissolved in the oil is about 2-6% based on the weight of the oil.

7. The isolated potentially regenerative cells of claim 2, wherein the oil comprises an oil selected from the group consisting of corn oil, peanut oil, cottonseed oil, rice bran oil, safflower oil, tea tree oil, pine nut oil, macadamia nut oil, camellia seed oil, rose hip oil, sesame oil, olive oil, soybean oil and combinations thereof.

8. The isolated potentially regenerative cells of claim 2, wherein the oil comprises sesame oil.

9. The isolated potentially regenerative cells of claim 2, wherein the sterol compound is selected from the group consisting of stigmasterol, campesterol, β

-sitosterol, chalinosterol, clionasterol, brassicasterol, α

-spinasterol, daucosterol, avenasterol, cycloartenol, desmosterol, and poriferasterol.

10. The isolated potentially regenerative cells of claim 2, wherein the sterol compound is a combination of stigmasterol, β

-sitosterol, and campesterol.

11. The isolated potentially regenerative cells of claim 2, wherein the sterol compound is a combination of stigmasterol and β

-sitosterol.

12. The isolated potentially regenerative cells of claim 2, wherein the sterol compound is a combination of brassicasterol and β

-sitosterol.

13. The isolated potentially regenerative cells of claim 2, wherein the sterol compound is a combination of brassicasterol, stigmasterol and β

-sitosterol.

14. The isolated potentially regenerative cells of claim 2, wherein the tissue culture medium further comprises wax that is dissolved in the fatty acid-containing oil and added to the water.

15. The isolated potentially regenerative cells of claim 14, wherein the concentration of the wax is about 1-20% by weight based on the weight of the oil.

16. The isolated potentially regenerative cells of claim 14, wherein the concentration of the wax is about 3-6% by weight based on the weight of the oil.

17. The isolated potentially regenerative cells of claim 14, wherein the wax is edible wax.

18. The isolated potentially regenerative cells of claim 14, wherein the wax is selected from the group consisting of beeswax, castorwax, glycowax, and carnaubawax, and beeswax.

19. The isolated potentially regenerative cells of claim 2, wherein the tissue culture medium further comprises baicalin dissolved in the oil.

20. The isolated potentially regenerative cells of claim 19, wherein the concentration of baicalin is about 0.001-2% by weight based on the weight of the oil.

21. The isolated potentially regenerative cells of claim 1, wherein the mammal is a rodent, a primate or a human.

22. The isolated potentially regenerative cells of claim 1, wherein the mammal is a fully developed adult human.

23. The isolated potentially regenerative cells of claim 1, wherein the mammal is alive.

24. The isolated potentially regenerative cells of claim 1, wherein the isolated potentially regenerative cells are from the brain, heart, liver, lung, intestine, stomach, kidney, bone marrow, or skin of the mammal, excluding embryonic stem cells and the blastocyst of the mammal.

25. The isolated potentially regenerative cells of claim 1, wherein the potentially regenerative cells contained in the isolated tissue cells or tissue are activated in the culture medium to continuously proliferate and differentiate for at least 5 days.

26. The isolated potentially regenerative cells of claim 1, wherein the potentially regenerative cells contained in the isolated tissue cells or tissue are activated in the culture medium to continuously proliferate and differentiate for at least 30 days.

27. The isolated potentially regenerative cells of claim 1, wherein the potentially regenerative cells contained in the isolated tissue cells or tissue are activated in the culture medium to continuously proliferate and differentiate for at least 50 days.

28. The isolated potentially regenerative cells of claim 1, wherein the physiological function of the tissue-organ includes the ability to produce biomolecules with activity selected from the group consisting of enzymatic activity, signaling and regulatory functions, and the ability to cause muscle contraction in response to electric current.