Measuring Clinically Relevant Endpoints in a Serum-Free, Three-Dimensional, Primary Cell Culture System of Human Osteoarthritic Articular Chondrocytes

Grace Bundens, Andrea Buckley, LaBraya Milton, Kathryn Behling, Sarah E Chmielewski, Ellen Cho, Xiomara Lozano-Torres, Abdulhafez Selim, Richard Lackman, Mindy George-Weinstein, Lawrence Miller, Marina D'Angelo

Research output: Contribution to journalArticlepeer-review

Abstract

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 × 10(6) 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.

Original languageAmerican English
JournalExperimental Cell Research
StatePublished - Jun 2 2017

Keywords

  • Collagen degradation
  • Human articular chondrocytes
  • Interleukin-1 beta
  • Osteoarthritis
  • Primary culture
  • Serum free cell culture

Disciplines

  • Medicine and Health Sciences

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