Generation of human immortalized chondrocytes from osteoarthritic and healthy cartilage: a new tool for cartilage pathophysiology studies

Bone & Joint Research

article

María Piñeiro-Ramil¹  Clara Sanjurjo-Rodríguez¹  Silvia Rodríguez-Fernández¹  Tamara Hermida-Gómez²  Francisco J. Blanco-García²  Isaac Fuentes-Boquete¹  Carlos Vaamonde-García²  Silvia Díaz-Prado¹ 

[1] Grupo de Investigación en Terapia Celular y Medicina Regenerativa, Universidade da Coruña ,(UDC), Instituto de Investigación Biomédica de A Coruña ,(INIBIC), Complexo Hospitalario Universitario de A Coruña;Centro de Investigacións Científicas Avanzadas ,(CICA), Universidade da Coruña

关键词: Articular chondrocytes;    Osteoarthritis;    Cell immortalization;    Inflammation;    chondrocytes;    cartilage tissues;    articular chondrocytes;    articular cartilage;    interleukin;    osteoarthritis (OA);    diarthrodial joints;    type I collagen;    cytokine;    primary OA;   

DOI  :  10.1302/2046-3758.121.BJR-2022-0207.R1

学科分类：骨科学

来源: British Editorial Society Of Bone And Joint Surgery

【 摘 要 】

AimsAfter a few passages of in vitro culture, primary human articular chondrocytes undergo senescence and loss of their phenotype. Most of the available chondrocyte cell lines have been obtained from cartilage tissues different from diarthrodial joints, and their utility for osteoarthritis (OA) research is reduced. Thus, the goal of this research was the development of immortalized chondrocyte cell lines proceeded from the articular cartilage of patients with and without OA.MethodsUsing telomerase reverse transcriptase (hTERT) and SV40 large T antigen (SV40LT), we transduced primary OA articular chondrocytes. Proliferative capacity, degree of senescence, and chondrocyte surface antigen expression in transduced chondrocytes were evaluated. In addition, the capacity of transduced chondrocytes to synthesize a tissue similar to cartilage and to respond to interleukin (IL)-1β was assessed.ResultsCoexpression of both transgenes (SV40 and hTERT) were observed in the nuclei of transduced chondrocytes. Generated chondrocyte cell lines showed a high proliferation capacity and less than 2% of senescent cells. These cell lines were able to form 3D aggregates analogous to those generated by primary articular chondrocytes, but were unsuccessful in synthesizing cartilage-like tissue when seeded on type I collagen sponges. However, generated chondrocyte cell lines maintained the potential to respond to IL-1β stimulation.ConclusionThrough SV40LT and hTERT transduction, we successfully immortalized chondrocytes. These immortalized chondrocytes were able to overcome senescence in vitro, but were incapable of synthesizing cartilage-like tissue under the experimental conditions. Nonetheless, these chondrocyte cell lines could be advantageous for OA investigation since, similarly to primary articular chondrocytes, they showed capacity to upregulate inflammatory mediators in response to the IL-1β cytokine.

【 授权许可】

CC BY-NC