Preparation method for therapeutic agent of bead-type chondrocyte
The present invention relates to a preparation method for a therapeutic agent of a bead-type chondrocyte and, more specifically, to: a preparation method for a therapeutic agent of a bead-type chondrocyte without a support, comprising the steps of: a) dispensing chondrocytes and/or cells having chondrogenic differentiation capability in a 96-well deep well plate having a V-shaped bottom; b) centrifuging the plate; c) three-dimensionally culturing the plate in an incubator; and d) recovering pellets from each well, thereby being capable of easily and stably preparing cartilage tissues of a uniform quality, in large quantities; and a therapeutic agent of a bead-type chondrocyte without a support, prepared by the method, having effectiveness to repair damage by simply being implanted into cartilage damage by injection.
1. A method for preparing a therapeutic agent of bead-type chondrocyte which comprises the following steps:
a) dispensing chondrocyte and/or cell having chondrogenic potential into a 96-well deep well plate having V-shaped bottom and well volume of 500 μl or more, with 0.5×10 5 cells to 2.0×10 5 cells/400 to 600 μl/well;
b) centrifuging the plate;
c) three-dimensionally culturing the plate in an incubator; and
d) collecting a pellet from each well.
2. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 1 , wherein the cell having chondrogenic potential in step (a) is selected from the group consisting of mesenchymal stem cell, embryonic stem cell and induced pluripotent stem cell.
3. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 2 , wherein the mesenchymal stem cell is adipose-derived, bone marrow-derived, umbilical cord-derived, umbilical cord blood-derived, placenta-derived, synovium-derived, periosteum-derived or perichondrium-derived cell.
4. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 1 , wherein said chondrocyte or cell is dispensed with 0.5×10 5 cells to 2.0×10 5 cells/400 μl/well in step (a).
5. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 1 , wherein said chondrocyte or cell is dispensed by the use of an apparatus selected from the group consisting of a multi-channel pipette, a multi-pipette, a microplate washer and a microplate dispenser in step (a).
6. The method for preparing a therapeutic agent of bead-type according to claim 1 , wherein the centrifugation is carried out at 500 to 3,000 rpm for 5 to 10 minutes in step (b).
7. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 1 , wherein the three-dimensional culture is carried out in a serum-free culture medium in step (c).
8. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 1 , wherein the three-dimensional culture is carried out for 3 days to 30 days in step (c).
9. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 1 , which further comprises a step of exchanging a culture medium in step (c).
10. The method for preparing a therapeutic agent of bead-type chondrocyte according to claim 9 , wherein the exchange of culture medium is carried out by the use of an apparatus selected from the group consisting of a multi-channel pipette, a multi-pipette, a microplate washer and a microplate dispenser.