Claims
- 1. A method for preparing Signal-plexes comprising:
a) harvesting animal tissue; b) lysing, homogenizing and filtering said tissue to produce extracts; c) fractionating said tissue to produce extracts and fractions thereof; and, d) testing the biological activity of said extracts and fractions thereof to select those which optimally induce differentiation.
- 2. The method of claim 1, wherein said tissue is selected from the group consisting of embryonic, fetal and post-natal tissue.
- 3. The method of claim 1, wherein said solids comprise cell membranes, nuclear membranes, nuclei and mitochondria.
- 4. A Signal-plex prepared by the method of claim 1.
- 5. A Signal-plex comprising biomolecules that are capable of inducing cells derived from animal tissue to differentiate and form tissue and organ primordia in vitro and are substantially free of nucleic acids, cell membranes, nuclear membranes, nuclei, mitochondria and microorganisms.
- 6. A method for differentiating cells to form tissue and organ primordia in vitro comprising:
a) providing cells derived from animal tissue; b) delivering said cells to a substrate; c) cultivating said cells in a culture medium with serum; d) providing Signal-plex(es); and, e) exposing said cells to said Signal-plex(es) for a period of 1 day to 14 months.
- 7. The method of claim 6, wherein said tissue is selected from the group consisting of embryonic, fetal and post-natal tissue.
- 8. The method of claim 6, wherein a subpopulation of cells in said tissue are progenitor cells and stem cells.
- 9. The method of claim 8, wherein said cells are fetal dermal fibroblasts harboring a subpopulation of said stem cells called Hu abba-cells.
- 10. The method of claim 8, wherein said cells are post-natal dermal fibroblasts from a person of any age harboring a subpopulation of said stem cells not named.
- 11. The method of claim 8, wherein said progenitor and stem cells in embryonic, fetal and post-natal tissues are induced to form when exposed to said Signal-plex(es).
- 12. The method of claim 6, wherein said substrate is selected from the group consisting of a cover glass, tissue culture plate, collagen fiber scaffold and collagen gel scaffold.
- 13. The method of claim 6, wherein said delivering comprises adding said cells to a scaffold.
- 14. The method of claim 11, wherein said scaffold is three-dimensional.
- 15. The method of claim 11, wherein said scaffold comprises at least one type of collagen.
- 16. The method of claim 13, wherein said scaffold is selected from the group consisting of a hydrated collagen fiber, collagen gel, collagen foam, freeze dried collagen and EBM.
- 17. The method of claim 6, wherein said cultivating does not comprise subculturing.
- 18. The method of claim 6, wherein said cultivating comprises renewing said Signal-plex(es) and said medium every 3 to 4 days.
- 19. The method of claim 6, wherein said cells exposed to Signal-plex(es) form aggregates of small cells that undergo morphogenesis to become nodules; and, cells derived from said nodules upon enzymatic dissociation of said nodules differentiate into new aggregates that form new nodules.
- 20. The method of claim 6, wherein said exposing comprises exposing said cells to one of said Signal-plex(es) at a time.
- 21. A cell(s) differentiated by the method of claim 6.
- 22. A cell(s) of claim 21, wherein said cell(s) is selected from the group consisting of endocrine and exocrine pancreas, liver, lung, kidney, heart, cartilage, vascular, tendon, ligament and bone cells.
- 23. A tissue primordium formed from a cell(s) of claim 21.
- 24. An organ primordium formed from a cell(s) of claim 21.
- 25. A cell(s) of claim 21, wherein said cell(s) is selected from the group consisting of endocrine and exocrine pancreas, liver, lung, kidney, heart, cartilage, vascular, tendon, ligament and bone cells.
- 26. A method for using Signal-plexes to treat a patient comprising delivering said cell(s) of claim 21 and tissue and organ primordium formed by said cell(s) to a patient, wherein said patient is selected from the group consisting of a human, an animal of high economic value, and an animal of high attachment value.
- 27. The method of claim 26, wherein said delivering comprises grafting said cell(s) and tissue and organ primordium formed by said cell(s) into said patient.
- 28. The method of claim 26, wherein said delivering comprises injecting said cell(s) and tissue and organ primordium formed by said cell(s) into said patient.
- 29. A method of using cells dissociated from aggregates of small cells for promoting cell divisions and scaled-up propagation of differentiated cells in vitro comprising:.
a) providing cells of claim 19; b) cutting out aggregates of small cells by using a device selected from the group consisting of a pair of fine scissors and cloning ring; c) dissociating said aggregates of small cells enzymatically into single cells to create a suspension of cells; d) diluting said suspension of cells; e) plating said cells at low density; f) culturing and passaging said cells in the presence of leukemia inhibitory factor (LIF) in a bioreactor to increase the numbers of said cells; and, g) exposing said cells to Signal-plex(es) after the removal of said LIF to form fully differentiated cells.
- 30. A differentiated cell(s) formed by the method of claim 29.
- 31. A tissue primordium formed by a differentiated cell(s) of claim 30.
- 32. An organ primordium formed by a differentiated cell(s) of claim 30.
- 33. A method for using differentiated cells for treating a patient by delivering a cell(s) of claim 36, and tissue and organ primordium formed by said cell(s) to a patient, wherein said patient is selected from the group consisting of a human, an animal of high economic value, and an animal of high attachment value.
- 34. The method of claim 33, wherein said delivering comprises grafting said cell(s) and tissue and organ primordium into said patient.
- 35. The method of claim 33, wherein said delivering comprises injecting said cell(s) and tissue and primordium into said patient.
- 36. A method for identifying stem cells in animal tissue comprising:
a) providing a cell(s) of claim 21; b) assaying said cells for tissue-specific properties.
- 37. The method of claim 36, wherein said assaying comprises performing a test selected from the group consisting of morphology, immunostaining, ELISA and RT-PCR.
- 38. A stem cell identified by the method of claim 36.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation-in-part of U.S. Ser. No. 09/901,765, filed Jul. 9, 2001, the entire contents of which are incorporated herein by reference.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60251125 |
Dec 2000 |
US |
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
09901765 |
Jul 2001 |
US |
Child |
10098158 |
Mar 2002 |
US |