Method for culturing mesenchymal stem cells
Disclosed is a method for culturing mesenchymal stem cells, comprising culturing mesenchymal stem cells in a medium containing calcium in a concentration of from 2.1 to 3.8 mM and magnesium in a concentration of from 1.0 to 3.0 mM under a hypoxic condition of 2 to 5% oxygen. The culturing method can increase the population of mesenchymal stem cells even with a small number of passages by improving mesenchymal stem cells in proliferative capacity and viability. In addition, the mesenchymal stem cells prepared by the culturing method are effectively used not only as a safe cell therapeutic agent due to their lacking immunogenicity, but also as a cartilage regenerating medicine owing to their excellent secretion of cytokines.
1. A method for forming bone and/or regenerating cartilage in a subject in need thereof, comprising administering a cell therapeutic agent comprising mesenchymal stem cells into the subject,
wherein said mesenchymal stem cells are obtained by a culture method comprising:
(a) culturing mesenchymal stem cells in a medium containing calcium in a concentration of from 2.7 to 3.8 mM under a hypoxic condition of 2 to 5% oxygen.
2. The method of claim 1 , wherein the mesenchymal stem cells of (a) are derived from umbilical cord blood, bone marrow, lipid, muscle, skin, amniotic fluid, umbilical cord, or teeth.
3. The method of claim 2 , wherein the medium is selected from the group consisting of a Dulbecco's modified eagle medium (DMEM), a minimal essential medium (MEM), an α-MEM, a McCoys 5A medium, an eagle's basal medium, a CMRL (Connaught Medical Research Laboratory) medium, a Glasgow MEM, a Ham's F-12 medium, an IMDM (Iscove's modified Dulbecco's medium), a Leibovitz's L-15 medium, an RPMI (Roswell Park Memorial Institute) 1640 medium, a medium 199, and a Hank's medium 199.
4. The method of claim 3 , wherein the medium comprises 5 to 30% of fetal bovine serum.
5. The method of claim 3 , wherein the medium does not comprise fetal bovine serum, but a serum replacement.
6. The method of claim 1 , wherein the medium is based on an α-MEM supplemented with 5 to 30% of fetal bovine serum (FBS), 0.3 to 2.0 mM of calcium, and 0.2 to 2.2 mM of magnesium.
7. The method of claim 1 , wherein the culture method further comprises sub-culturing the mesenchymal stem cells obtained in (a) in the same condition as in claim 1 .
8. The method of claim 1 , wherein the medium of (a) further comprises magnesium in a concentration of from 1.0 to 3.0 mM.
9. The method of claim 1 , wherein the medium of (a) comprises calcium in a concentration of from 3.3 to 3.8 mM.