Mammary artery derived cells and methods of use in tissue repair and regeneration
View Patent ↗An isolated mammalian internal mammary artery-derived cell is disclosed. Furthermore, methods of isolating the mammalian internal mammary artery-derived cell are disclosed. The cell is useful in tissue engineering technologies, specifically in vascular tissue engineering.
1. An in vitro cell culture comprising:
A culture vessel;
A culture medium comprising internal mammary artery-derived cell growth medium; and
A homogenous population of cells capable of self-renewal and expansion in culture derived from a mammalian internal mammary artery,
Wherein said cells are positive for the expression of cell-surface marker HLA-1 and negative for expression of each one of cell-surface markers CD 10, CD31, CD34, CD45, CD133, CD141 and KDR/Flk-1, and
Wherein said cells are plated in the culture vessel with the culture medium.
2. The cell culture of claim 1 , wherein the cells are capable of differentiation into adipocytes, cardiomyocytes, and endothelial cells.
3. The cell culture of claim 1 wherein the culture vessel is coated.
4. The cell culture of claim 3 wherein the culture vessel is coated with collagen.
5. The cell culture of claim 1 wherein the cells are additionally positive for expression of CD29, CD44, CD73, CD166, and additionally negative for CD15, CD23, CD24, CD62p, CD80, CD86, CD104, CD117, CD138, CD146, VE-Cadherin, and HLA-2.
6. An in vitro homogenous cell culture produced by a method comprising the steps of:
providing a mammalian internal mammary artery,
providing an enzyme mixture wherein the enzyme mixture is comprised of digestion enzymes selected from the group consisting of a metalloprotease enzyme, a neutral protease enzyme, a mucolytic enzyme, and combinations thereof,
incubating the entire mammalian internal mammary artery in the enzyme mixture for a time sufficient to remove the intimal layer and to remove external debris from the artery and provide a partially digested artery,
transferring the partially digested artery into a fresh enzyme mixture and continue digestion for a time sufficient to provide a digested material,
isolating and culturing the cells from the digested material to obtain a homogenous population of cells, wherein the cells are positive for the expression of cell-surface marker HLA-1 and negative for expression of CD34,
suspending the cells in a culture medium, and
plating the cells in a culture vessel.
7. The cell culture of claim 6 wherein the enzyme mixture is comprised of collagenase and dispase.
8. The cell culture of claim 6 wherein the time to remove the intimal layer and to remove external debris from the artery is about 60 minutes.
9. The cell culture of claim 6 wherein the time sufficient to provide digested material is in the range of about 30 minutes to about 60 minutes.
10. The cell culture of claim 6 wherein the time sufficient to saturate the enzyme mixture with digested material is about 60 minutes.
11. An implant comprising a population of isolated internal mammary artery-derived cells in a biocompatible matrix, wherein the population of isolated internal mammary artery-derived cells are capable of self-renewal and expansion in culture and are positive for the expression of cell-surface marker HLA-1 and negative for expression of each one of cell-surface markers CD 10, CD31, CD34, CD45, CD133, CD141 and KDR/F1k-1.
12. The implant of claim 11 wherein the population of isolated internal mammary artery-derived cells are additionally positive for expression of CD29, CD44, CD73, CD166, and additionally negative for CD15, CD23, CD24, CD62p, CD80, CD86, CD104, CD117, CD138, CD146, VE-Cadherin, and HLA-2.
13. The implant of claim 11 wherein the population of isolated internal mammary artery-derived cells are capable of differentiation into adipocytes, cardiomyocytes, and endothelial cells.