Method for preparing pellets of chondrocytes from human induced pluripotent stem cells, and use thereof
A method for preparing chondrocyte pellets including: (a) culturing human induced pluripotent stem cells so as to form and obtain embryoid bodies; (b) inducing the embryonic bodies obtained in step (a) to become outgrowth cells and isolating same; and (c) culturing the outgrowth cells isolated in step (b) in a pellet form. A pharmaceutical composition including the chondrocyte pellets prepared by the preparation method; or a method for preventing or treating arthritis, including administering the chondrocyte pellets prepared by the method to an arthritis patient. Chondrocyte pellets of the present invention have a remarkably high rate of differentiation into chondrocytes, uniform size, and a homogeneous differentiation degree, and a pharmaceutical composition, including the pellets, has excellent cartilage regeneration effects at the site of cartilage damage, and is in injectable form, which does not require surgery, such that pain is relieved and a continuous arthritis treatment is provided.
1. A method of preparing an injectable chondrocyte pellet comprising differentiated hyaline chondrocytes, the method comprising:
(a) culturing human induced pluripotent stem cells (hiPSCs) derived from cord blood mononuclear cells to form and obtain an embryoid body;
(b) inducing the embryoid body obtained in Step (a) into outgrowth cells and isolating the outgrowth cells;
(c) centrifuging 1,500 to 2,500 of the outgrowth cells isolated in Step (b) at a speed of 1,600 to 2,000 rpm to form a chondrocyte pellet; and
(d) culturing the chondrocyte pellet formed in step (c) in a serum-free medium comprising a human bone morphogenetic protein (BMP) and a transforming growth factor-beta (TGF-β),
wherein the chondrocyte pellet in step (c) has a diameter of 200 to 300 μm.
2. The method of claim 1 , wherein the culture in Step (a) is adherent culture.
3. The method of claim 1 , wherein the induction in Step (b) is performed on a gelatin-coated plate.
4. A method of treating an arthritis, comprising:
administering the chondrocyte pellet prepared by the method of claim 1 to an arthritis patient via intra-articular injection.
5. The method of claim 4 , wherein the administration is performed using a syringe.
6. The method of claim 4 , wherein the arthritis is any one or more selected from osteoarthritis, rheumatoid arthritis, psoriatic arthritis, septic arthritis, osteochondritis dissecans, arthritis due to joint and ligament damage, and arthritis due to meniscus injury.