TY - JOUR
T1 - Measuring clinically relevant endpoints in a serum-free, three-dimensional, primary cell culture system of human osteoarthritic articular chondrocytes
AU - Bundens, Grace
AU - Buckley, Andrea
AU - Milton, La Braya
AU - Behling, Kathryn
AU - Chmielewski, Sarah
AU - Cho, Ellen
AU - Lozano-Torres, Xiomara
AU - Selim, Abdulhafez
AU - Lackman, Richard
AU - George-Weinstein, Mindy
AU - Miller, Lawrence
AU - D'Angelo, Marina
N1 - Publisher Copyright:
© 2017 Elsevier Inc.
PY - 2017/8/15
Y1 - 2017/8/15
N2 - Osteoarthritis (OA) is characterized by degeneration of articular cartilage within the joint, inflammation and pain. The purpose of this study was to develop a primary, serum free cell culture system of human osteoarthritic articular chondrocytes (HOACs) with which to study manifestations of the disease process. Joint tissues were obtained from OA patients undergoing total knee arthroplasty (TKA). HOACs isolated from the femoral condyles and tibial plateau of the same side were combined, plated in three-dimensional, alginate beads and cultured for five days in serum, hormone and protein free medium. More living cells were obtained from the femoral condyles than the tibial plateau. The optimal plating density was 2.5 × 106 cells/ml of alginate. The amounts of DNA, RNA, proteoglycans and total collagen were similar in cultures prepared from the sides of least and greatest pathology. More type 1 than type 2 collagen was detected in the medium on days 2 and 5. A greater percentage of type 1 than type 2 collagen was degraded. The inflammatory cytokine interleukin-1 beta was present in the medium and alginate associated matrix. Although variation in the metabolic profiles between subjects was observed, HOACs from all patients continued to reflect the OA phenotype for five days in culture. This serum free, three-dimensional primary culture system of HOACs provides a platform with which to measure clinically relevant endpoints of OA and screen potential disease modifying OA therapeutics.
AB - Osteoarthritis (OA) is characterized by degeneration of articular cartilage within the joint, inflammation and pain. The purpose of this study was to develop a primary, serum free cell culture system of human osteoarthritic articular chondrocytes (HOACs) with which to study manifestations of the disease process. Joint tissues were obtained from OA patients undergoing total knee arthroplasty (TKA). HOACs isolated from the femoral condyles and tibial plateau of the same side were combined, plated in three-dimensional, alginate beads and cultured for five days in serum, hormone and protein free medium. More living cells were obtained from the femoral condyles than the tibial plateau. The optimal plating density was 2.5 × 106 cells/ml of alginate. The amounts of DNA, RNA, proteoglycans and total collagen were similar in cultures prepared from the sides of least and greatest pathology. More type 1 than type 2 collagen was detected in the medium on days 2 and 5. A greater percentage of type 1 than type 2 collagen was degraded. The inflammatory cytokine interleukin-1 beta was present in the medium and alginate associated matrix. Although variation in the metabolic profiles between subjects was observed, HOACs from all patients continued to reflect the OA phenotype for five days in culture. This serum free, three-dimensional primary culture system of HOACs provides a platform with which to measure clinically relevant endpoints of OA and screen potential disease modifying OA therapeutics.
UR - http://www.scopus.com/inward/record.url?scp=85020227699&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85020227699&partnerID=8YFLogxK
U2 - 10.1016/j.yexcr.2017.06.001
DO - 10.1016/j.yexcr.2017.06.001
M3 - Article
C2 - 28583763
AN - SCOPUS:85020227699
SN - 0014-4827
VL - 357
SP - 310
EP - 319
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 2
ER -