P072 - Propofol as Anaesthetic – Does it Impair Manufacturing of Autologous Chondrocyte Implantation Products?
- A. Schubert (Teltow, DE)
- A. Schubert (Teltow, DE)
- G. Roel (Teltow, DE)
- P. Giesemann (Teltow, DE)
- C. Kaps (Teltow, DE)
- L. Vonk (Teltow, DE)
- A. Hoburg (Berlin, DE)
Abstract
Purpose
In clinical practice, human autologous chondrocytes are used for implantation to treat cartilage defects. For chondrocyte culture, autologous serum is often used as supplement in the culture medium. It is under discussion, whether drawing the blood during anaesthesia for the cartilage biopsy harvest impairs the manufacturing of the autologous cell implant. Since many clinicians use the anaesthetic propofol, we investigated a possible impact of propofol on cell’s performance during cell expansion and 3D culture.
Methods and Materials
Blood was obtained from 10 donors (male/female 4/6, 30-49 years) before and after propofol administration (1% in soy oil, 2,3-4,2 mg/kg/h, 4-21 minutes after administration) to generate serum without and with propofol. Human primary chondrocytes were isolated from 3 donors (male/female 2/1, 56-76 years). Cell expansion and 3D spheroid formation were performed using both normal serum and serum with propofol (n=10). Cell adhesion was evaluated 5 days after seeding and gene expression of cartilage-matrix proteoglycan aggrecan (ACAN) was measured in chondrocyte spheroids after two weeks of culture.
Results
Serum with propofol was cloudy in 8/10 batches. Chondrocytes were expandable and 3D spheroids were formed using both serum with and without propofol. However, the propofol group showed lower cell adhesion in 5/10 batches (Figure 1A) and 3 of these showed less population doublings in P0 (Figure 1B) and thus additional passaging was necessary to achieve similar cell yields. ACAN gene expression was decreased in 3 spheroid cultures in the propofol group (Figure 2).
Conclusion
In presence of propofol, chondrocytes could still grow in 2D culture and assembled as spheroids in 3D culture. However, serum enriched with propofol may affect initial cell adhesion, to extend cell culture time, or change cartilage-specific gene expression in some chondrocyte cultures. Therefore, it is not recommended to draw the blood for autologous serum applications from the patient under anaesthesia with propofol.