Poster Culture Models

P010 - The Differentiation of Human Chondrocytes in Scaffold-free 3D Tissue Culture is Independent of the Utilized Serum Type

Presentation Topic
Culture Models
Date
13.04.2022
Lecture Time
09:30 - 09:30
Room
Exhibition Foyer
Session Name
7.3 - Poster Viewing / Coffee Break / Exhibition
Session Type
Poster Session
Speaker
  • A. Ecke (Senftenberg, DE)
Authors
  • A. Ecke (Senftenberg, DE)
  • J. Scholka (Senftenberg, DE)
  • E. Hentschel (Senftenberg, DE)
  • U. Anderer (Senftenberg, DE)
Disclosure
No Significant Commercial Relationship

Abstract

Purpose

In vitro cell culture and tissue engineering techniques often rely on animal sera such as foetal bovine serum (FBS) or on human serum (HS). However, reproducibility of experiments and applicability of the results on in vivo situations is limited. Furthermore, there are ethical concerns (1) with the production of FBS via killing calves in the womb, and (2) the use of blood donations for the production of HS for research. Hence, the aim of this study was to compare the differentiation degree of cartilage-like microtissues cultivated with species specific sera and a chemically defined serum (CDS).

Methods and Materials

Human primary chondrocytes isolated from knee joints were expanded in medium containing HS. Scaffold-free microtissues were generated and cultivated in cell-repellent plates using three different sera: HS, FBS and CDS. After two and four weeks, the macroscopic appearance (reflected-light microscopy) was documented and the size was determined. The differentiation degree was evaluated via histology to visualize typical glycosaminoglycans (Safranin O, Alcian blue) and immunohistochemistry (IH) to detect cartilage-specific markers (collagen type II, proteoglycans, Sox9) and collagen type I on cryosections.

Results

All microtissues had a whitish colour both after two and four weeks cultivation. Cartilage-like microtissues cultivated with HS or FBS were similar in size, whereas those cultivated with CDS were more than 10% bigger. Already after two weeks, cartilage-like microtissues of all tested sera showed an expression of cartilage-specific markers, which increased after four weeks. Collagen type I was always weakly expressed. The expression level of all tested markers was similar regardless of serum type.

Conclusion

This study suggests that the differentiation degree of cartilage-like microtissues in cell-repellent plates is not influenced by the serum type. Hence, CDS can replace species specific sera to reduce ethical concerns and batch-to-batch variations. The applicability of CDS to the expansion of chondrocytes remains to be determined.

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