Chondrocytes expressing intracellular collagen type II enter the cell cycle and co-express collagen type I in monolayer culture.

Tekari, Adel; Luginbuehl, Reto; Hofstetter, Wilhelm; Egli, Rainer (2014). Chondrocytes expressing intracellular collagen type II enter the cell cycle and co-express collagen type I in monolayer culture. Journal of orthopaedic research, 32(11), pp. 1503-1511. Wiley 10.1002/jor.22690

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For autologous chondrocyte transplantation, articular chondrocytes are harvested from cartilage tissue and expanded in vitro in monolayer culture. We aimed to characterize with a cellular resolution the synthesis of collagen type II (COL2) and collagen type I (COL1) during expansion in order to further understand why these cells lose the potential to form cartilage tissue when re-introduced into a microenvironment that supports chondrogenesis. During expansion for six passages, levels of transcripts encoding COL2 decreased to <0.1%, whereas transcript levels encoding COL1 increased 370-fold as compared to primary chondrocytes. Flow cytometry for intracellular proteins revealed that chondrocytes acquired a COL2/COL1-double positive phenotype during expansion, and the COL2 positive cells were able to enter the cell cycle. While the fraction of COL2 positive cells decreased from 70% to <2% in primary chondrocytes to passage six cells, the fraction of COL1 positive cells increased from <1% to >95%. In parallel to the decrease of the fraction of COL2 positive cells, the cells' potential to form cartilage-like tissue in pellet cultures steadily decreased. Intracellular staining for COL2 enables for characterization of chondrocyte lineage cells in more detail with a cellular resolution, and it may allow predicting the effectiveness of expanded chondrocytes to form cartilage-like tissue.

Item Type:

Journal Article (Original Article)

Division/Institute:

04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR)
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DBMR Forschung Mu35 > Forschungsgruppe Knochenbiologie & Orthopädische Forschung
04 Faculty of Medicine > Pre-clinic Human Medicine > BioMedical Research (DBMR) > DBMR Forschung Mu35 > Forschungsgruppe Knochenbiologie & Orthopädische Forschung

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Tekari, Adel; Luginbuehl, Reto; Hofstetter, Wilhelm and Egli, Rainer

Subjects:

600 Technology > 610 Medicine & health

ISSN:

0736-0266

Publisher:

Wiley

Language:

English

Submitter:

Verena de Serra Frazao-Bill

Date Deposited:

23 Apr 2015 16:04

Last Modified:

02 Nov 2015 11:01

Publisher DOI:

10.1002/jor.22690

PubMed ID:

25043137

Uncontrolled Keywords:

bromodeoxyuridine; chondrocyte; collagen type II; flow cytometry; monolayer culture

BORIS DOI:

10.7892/boris.64660

URI:

https://boris.unibe.ch/id/eprint/64660

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