FLOW CYTOMETRIC ANALYSIS OF NORMAL AND OSTEOARTHRITIC
CHONDROCYTES

DLUGOŠOVÁ, Slavomíra, Zuzana KAŇOVSKÁ and Irena KOUTNÁ. FLOW CYTOMETRIC ANALYSIS OF NORMAL AND OSTEOARTHRITIC CHONDROCYTES. In 12th International Conference Analytical Cytometry, Prague, Czech Republic. 2023.

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TY  - CONF
ID  - 2380517
AU  - Dlugošová, Slavomíra - Kaňovská, Zuzana - Koutná, Irena
PY  - 2023
TI  - FLOW CYTOMETRIC ANALYSIS OF NORMAL AND OSTEOARTHRITIC CHONDROCYTES
KW  - Osteoarthritis
KW  - flow cytomeric analysis
N2  - Osteoarthritis (OA) is the most common degenerative joint disease characterized by increased degradation of cartilage tissue in the joint due to overexpression of proteolytic enzymes degrading extracellular matrix (ECM). OA is characterized by cartilage degradation, osteophyte formation, and stiffening of joints. Due to the inability of resident chondrocytes to regenerate the ECM with the same properties as it was formed during development, the probability of spontaneous regeneration is very low. Treatment is aimed at alleviating symptoms such as pain and swelling. The last option is a total replacement of the joint. Regenerative medicine using cells as therapeutics offers a large hope in treating OA. The main problem and challenge of advanced therapy medicinal product (ATMP) based on cultured human articular chondrocytes (HACs) is that chondrocytes transferred into monolayer culture, lose their typical properties – round morphology changes to fibroblast-like spindle shape, and cartilaginous protein synthesis decreases. This phenomenon is termed dedifferentiation. Our working hypothesis was that modification of growth media can modify chondrocytes’ properties, ideally increasing the synthesis of ECM, mainly collagen type II (COL2). We needed to identify surface markers correlated with the differentiation status of monolayer expanded HACs and to define the degree of dedifferentiation. We selected 3 surface markers that should determine the state of the cell (CD44, CD54, CD90). Our results indicate that increased expression of CD54 is therefore associated with a response to inflammatory stimulation. In the absence of an inflammatory reaction, the cellular distribution of this marker is reduced, in healthy cartilage the expression of this marker was not detected at all. The high level of expression of the markers CD44 and CD90 is directly connected with the good chondrogenic capacity of the cells and the ability to produce a matrix with a high content of COL2.
ER  -

Basic information
Original name	FLOW CYTOMETRIC ANALYSIS OF NORMAL AND OSTEOARTHRITIC CHONDROCYTES
Authors	DLUGOŠOVÁ, Slavomíra (703 Slovakia, belonging to the institution), Zuzana KAŇOVSKÁ (203 Czech Republic) and Irena KOUTNÁ (203 Czech Republic, belonging to the institution).
Edition	12th International Conference Analytical Cytometry, Prague, Czech Republic, 2023.

Other information
Original language	English
Type of outcome	Conference abstract
Field of Study	10601 Cell biology
Country of publisher	Czech Republic
Confidentiality degree	is not subject to a state or trade secret
Organization unit	Faculty of Medicine
Keywords in English	Osteoarthritis; flow cytomeric analysis
Tags	International impact
Changed by	Changed by: Mgr. Tereza Miškechová, učo 341652. Changed: 19/7/2024 11:46.

Abstract

Osteoarthritis (OA) is the most common degenerative joint disease characterized by increased degradation of cartilage tissue in the joint due to overexpression of proteolytic enzymes degrading extracellular matrix (ECM). OA is characterized by cartilage degradation, osteophyte formation, and stiffening of joints. Due to the inability of resident chondrocytes to regenerate the ECM with the same properties as it was formed during development, the probability of spontaneous regeneration is very low. Treatment is aimed at alleviating symptoms such as pain and swelling. The last option is a total replacement of the joint. Regenerative medicine using cells as therapeutics offers a large hope in treating OA. The main problem and challenge of advanced therapy medicinal product (ATMP) based on cultured human articular chondrocytes (HACs) is that chondrocytes transferred into monolayer culture, lose their typical properties – round morphology changes to fibroblast-like spindle shape, and cartilaginous protein synthesis decreases. This phenomenon is termed dedifferentiation. Our working hypothesis was that modification of growth media can modify chondrocytes’ properties, ideally increasing the synthesis of ECM, mainly collagen type II (COL2). We needed to identify surface markers correlated with the differentiation status of monolayer expanded HACs and to define the degree of dedifferentiation. We selected 3 surface markers that should determine the state of the cell (CD44, CD54, CD90). Our results indicate that increased expression of CD54 is therefore associated with a response to inflammatory stimulation. In the absence of an inflammatory reaction, the cellular distribution of this marker is reduced, in healthy cartilage the expression of this marker was not detected at all. The high level of expression of the markers CD44 and CD90 is directly connected with the good chondrogenic capacity of the cells and the ability to produce a matrix with a high content of COL2.

Links
LM2023049, research and development project	Name: Český národní uzel Evropské sítě infrastruktur klinického výzkumu
LM2023049, research and development project	Investor: Ministry of Education, Youth and Sports of the CR

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FLOW CYTOMETRIC ANALYSIS OF NORMAL AND OSTEOARTHRITIC CHONDROCYTES

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