Understanding factors affecting human chondrocyte culturing: an experimental study.

Cell and Tissue Banking
Pelin KilicAcelya Yilmazer

Abstract

Over the years, surgical strategies have been developed in hope of full regeneration of the injured cartilage. In our study, we aimed to develop an optimized chondrocyte culture isolation technique as an active ingredient of a standardized autologous chondrocte implantation product, which is able to maintain the phenotype along with the molecular features of the cartilage. We compared different enzymes, which suggested optimal performance with collagenase type II at 5 mg/ml concentration. Thereafter, we observed that COL2 and GAG expression is substantially reduced with passaging. There was a need to omit passaging to reach the optimal isolation method. We then tested various growth factors and media in order to maintain the natural character of chondrocytes. Our study also suggested the highest COL2 and GAG expressions with the highest recovery in the presence of Advanced DMEM. Autologous chondrocyte implantation manufacturing approval was recently received from the national competent authority, making it possible to utilize the process engineering protocol developed with this study at our Tissue and Cell Manufacturing Center as a part of the autologous chondrocyte implantation manufacturing standard operation procedure (SOP).

References

Feb 28, 2008·Journal of Bone and Mineral Metabolism·Shigeyuki WakitaniKunio Takaoka
Aug 21, 2008·Nature Protocols·Marjolaine GossetClaire Jacques
Jul 24, 2012·The Journal of Surgical Research·Adelola O OseniAlexander M Seifalian
May 1, 2013·Journal of Biomedical Materials Research. Part a·Wei ShuiSherwin H Ho
Jun 12, 2014·Tissue Engineering. Part C, Methods·Ting Ting LauDong-An Wang
May 14, 2016·Biomaterials·Brian J HuangKyriacos A Athanasiou
Apr 25, 2018·Transfusion and Apheresis Science : Official Journal of the World Apheresis Association : Official Journal of the European Society for Haemapheresis·Erden AtillaGunhan Gurman

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Methods Mentioned

BETA
biopsy
PCR
light microscopy
flow cytometry

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Excel
GraphPad

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