Tissue engineered cartilage: Utilization of autologous serum and serum-free media for chondrocyte culture

The main focus of this study is to examine the possible use of autologous serum and serum free media in chondrocyte growth for tissue engineered cartilage. They focus on the the different rates of proliferation of chondrocytes between the media with autologous serum, serum free media and the media with standard fetal bovine serum (FBS).

Chondrocytes were first isolated by taking auricular cartilage from pigs and fragmenting the sample. The sample was then washed, digested with 0.3% collagenase II, and filtered. The filtrate was taken and cell pelleted for use in the experiment. The chondrocytes were plated and the sample was placed into three different flasks with equal concentrations. The chondrocytes were subjected to three different media: Ham F12 culture media with 10% FBS, Ham F12 culture media with 10% autologous serum and Ham F12 culture media with growth factors. The culture was changed twice a week with morphology examined using light microscopy. The cells were trypsinized at days 3, 6, 9, and 12 with 0.25% Trysin/EDTA and counted with a hematocytometer.

The results showed that while the cells in the media with FBS grew faster initially, the cells in the serum-free media eventually caught up to the same cell count while the cells in the media with autologous serum grew at a slower rate. These findings prove that it is possible for chondrocyte expansion using media with autologous serum and serum-free media. The potential of these results is that the use of different media could reduce possible immunological reactions or vector infections from the FBS.

I chose this paper due to the similarities with our current labs involving Rex and 3T3 cells. In our lab we determined that the calf serum we were using was possibly influencing our results by containing some proteins that were analyzed in our quantification of protein. By using the serum-free media we could have nullified the serum's effects. Also, this directly applies to our goals of TE cartilage. Lastly, I feel that this paper illustrates various techniques we learned in class such as passaging and cell counting.