A novel in vitro assay to study chondrocyte-to-osteoblast transdifferentiation.

Animals Cell Differentiation Cell Transdifferentiation Cells, Cultured Chondrocytes / metabolism Chondrogenesis / genetics Mice Osteoblasts Osteogenesis
Cartilage to bone transformation Chondrocyte Endochondral ossification Fracture healing In vitro assay Transdifferentiation

Journal

Endocrine
ISSN: 1559-0100
Titre abrégé: Endocrine
Pays: United States
ID NLM: 9434444

Informations de publication

Date de publication:
Jan 2022
Historique:
received: 05 05 2021
accepted: 14 08 2021
pubmed: 17 9 2021
medline: 23 3 2022
entrez: 16 9 2021
Statut: ppublish

Résumé

Endochondral ossification, which involves transdifferentiation of chondrocytes into osteoblasts, is an important process involved in the development and postnatal growth of most vertebrate bones as well as in bone fracture healing. To study the basic molecular mechanisms of this process, a robust and easy-to-use in vitro model is desirable. Therefore, we aimed to develop a standardized in vitro assay for the transdifferentiation of chondrogenic cells towards the osteogenic lineage. Murine chondrogenic ATDC5 cells were differentiated into the chondrogenic lineage for seven days and subsequently differentiated towards the osteogenic direction. Gene expression analysis of pluripotency, as well as chondrogenic and osteogenic markers, cell-matrix staining, and immunofluorescent staining, were performed to assess the differentiation. In addition, the effects of Wnt3a and lipopolysaccharides (LPS) on the transdifferentiation were tested by their addition to the osteogenic differentiation medium. Following osteogenic differentiation, chondrogenically pe-differentiated cells displayed the expression of pluripotency and osteogenic marker genes as well as alkaline phosphatase activity and a mineralized matrix. Co-expression of Col2a1 and Col1a1 after one day of osteogenic differentiation indicated that osteogenic cells had differentiated from chondrogenic cells. Wnt3a increased and LPS decreased transdifferentiation towards the osteogenic lineage. We successfully established a rapid, standardized in vitro assay for the transdifferentiation of chondrogenic cells into osteogenic cells, which is suitable for testing the effects of different compounds on this cellular process.

Identifiants

DOI: 10.1007/s12020-021-02853-4 PMID: 34529238 PMC: PMC8763722
pubmed: 34529238
doi: 10.1007/s12020-021-02853-4
pii: 10.1007/s12020-021-02853-4
pmc: PMC8763722
doi:

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

266-275

Informations de copyright

© 2021. The Author(s).

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Auteurs

Miriam E A Tschaffon (MEA)

Institute of Orthopedic Research and Biomechanics, University Medical Center Ulm, Ulm, Germany.
ORCID: 0000-0002-6409-9340

Stefan O Reber (SO)

Laboratory for Molecular Psychosomatics, Department of Psychosomatic Medicine and Psychotherapy, University of Ulm, Ulm, Germany.

Astrid Schoppa (A)

Institute of Orthopedic Research and Biomechanics, University Medical Center Ulm, Ulm, Germany.

Sayantan Nandi (S)

Institute of Comparative Molecular Endocrinology, University of Ulm, Ulm, Germany.

Ion C Cirstea (IC)

Institute of Comparative Molecular Endocrinology, University of Ulm, Ulm, Germany.

Attila Aszodi (A)

Laboratory of Experimental Surgery and Regenerative Medicine, Clinic for General, Trauma and Reconstructive Surgery, Klinikum der Universität München, Martinsried, Germany.

Anita Ignatius (A)

Institute of Orthopedic Research and Biomechanics, University Medical Center Ulm, Ulm, Germany.

Melanie Haffner-Luntzer (M)

Institute of Orthopedic Research and Biomechanics, University Medical Center Ulm, Ulm, Germany. melanie.haffner-luntzer@uni-ulm.de.

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Classifications MeSH

questionsmedicales.fr › Eucaryotes › Animaux › A novel in vitro assay to study...
questionsmedicales.fr › Phénomènes physiologiques cellulaires › Différenciation cellulaire › A novel in vitro assay to study...
questionsmedicales.fr › Phénomènes physiologiques cellulaires › Transdifférenciation cellulaire › A novel in vitro assay to study...
questionsmedicales.fr › Cellules › Cellules cultivées › A novel in vitro assay to study...
questionsmedicales.fr › Cellules › Cellules du tissu conjonctif › Chondrocytes › A novel in vitro assay to study...
questionsmedicales.fr › Phénomènes physiologiques de l'appareil locomoteur et du système nerveux › Phénomènes physiologiques du système locomoteur › Développement locomoteur › Chondrogenèse › A novel in vitro assay to study...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Rodentia › Muridae › Murinae › Souris › A novel in vitro assay to study...
questionsmedicales.fr › Cellules › Cellules du tissu conjonctif › Ostéoblastes › A novel in vitro assay to study...
questionsmedicales.fr › Phénomènes physiologiques de l'appareil locomoteur et du système nerveux › Phénomènes physiologiques du système locomoteur › Développement locomoteur › Biominéralisation › Développement osseux › Ostéogenèse › A novel in vitro assay to study...