Stem cells and matrix from cord tissue
Methods for isolating and culturing stem cells, making tissue matrix, making matrix infused with stem cells, and methods of stem cell therapy are provided.
1. A method, comprising:
a) obtaining a first umbilical cord tissue from a first newborn animal;
b) cleaning said first umbilical cord tissue with a sterilizing agent;
c) mechanically dissociating said first umbilical cord tissue to isolate mixed cells from said first umbilical cord tissue without separating vessels from the rest of the cord tissue, wherein said cells are a mixture of differentiated cells, stem cells and progenitor cells;
d) obtaining a second umbilical cord tissue from said first newborn animal or from a second newborn animal;
e) cutting said second umbilical cord tissue to a desired shape without separating vessels from the rest of the cord tissue and treating said second umbilical cord tissue to produce a decellularized tissue;
f) culturing said mixed cells together with said decellularized tissue such that said decellularized tissue provides a 3D scaffold for 3D culturing of said mixed cells, wherein said 3D culturing is in a 0.1% to 5% oxygen environment at 37° C. in a medium supplemented with serum, plasma or platelet lysate from said first newborn animal.
2. The method of claim 1 , whereby said first umbilical cord tissue and said second umbilical cord tissue are from the same newborn animal.
3. The method of claim 1 , wherein said dissociating step c) is by slicing or dicing said tissue with a blade.
4. The method of claim 1 , wherein step e) is non-enzymatic.
5. The method of claim 1 , where said 3D culturing step is performed in a humidified carbon dioxide cell culture chamber.
6. The method of claim 1 , where said decellularized tissue and said mixed cells are stored together or separately at temperatures between −80° C. and −196° C. before use in said culturing step f).
7. The method of claim 1 , where said decellularized tissue is lyophilized and stored at −80° C. before use in said culturing step f).
8. The method of claim 7 , where said lyophilized decellularized tissue is thawed in phosphate buffer solution or water.
9. The method of claim 7 , where said decellularized tissue is lyophilized and stored at room temperature before use in said 3D culturing step f).
10. The method of claim 9 , where said lyophilized tissue matrix is reconstituted with water or phosphate buffer solution.
11. A method of producing stem cells, comprising:
a) obtaining a first piece of umbilical cord from a newborn animal;
b) cleaning said first piece of umbilical cord with a sterilizing agent;
c) mechanically dissociating said first piece of umbilical cord to isolate mixed cells from said first piece of umbilical cord without separating vessels from the rest of the umbilical cord, wherein said mixed cells are a mixture of differentiated cells, stem cells and progenitor cells;
d) obtaining a second piece of umbilical cord from said newborn animal;
e) cutting said second piece of umbilical cord into a desired shape without separating vessels from the rest of the umbilical cord and treating said second piece of umbilical cord to produce a decellularized tissue;
f) culturing said mixed cells together with said decellularized tissue such that said decellularized tissue provides a 3D scaffold for 3D culturing of said cells, wherein said 3D culturing is in a 0.1% to 5% oxygen environment at 37° C. in a medium supplemented with serum, plasma or platelet lysate from cord blood from said newborn animal.