Comparison of media and sera used for in vitro culture of equine superficial digital flexor tendon explants.

The research study examines how different commercial nutrient mediums and sera influence protein synthesis and cell density maintenance in equine superficial digital flexor tendon tissue cultures.

Methods and Experimentation

To undertake this investigation, the researchers used multiple commercially available nutrient media and sera. They chose eight healthy horses aged between 2 to 4 years for the study. They experimented with the various nutrient media, including Dulbecco’s Modified Eagle’s Medium (DMEM), Ham’s F12 nutrient mixture, RPMI 1640 medium, Minimum Essential Medium with Earle’s salts, Minimum Essential Medium with Hanks’ salts etc, with 10% fetal bovine serum (FBS).

Next, they evaluated the effect of FBS, fetal horse serum, and donor horse serum, each at concentrations of 5%, 10%, and 15%, in RPMI 1640 medium. All these specimens were cultured in roller bottles at a temperature of 37 degrees Celsius and aerated each day using a gas mixture of 50% Oxygen, 45% Nitrogen and 5% Carbon dioxide.

Measurements and Outcomes

To evaluation protein synthesis, the researchers measured the rate of [3H]-proline incorporation on the 13th and 28th days of the experiment. This particular approach served as an indicator of total protein and collagen synthesis. Further, the research team also assessed the matrix cellular density of the cultured explants at days 14 and 28 using a computerized image analysis system.

The experiment found that proline incorporation was at its peak in the Ham’s F12 nutrient mixture and RPMI 1640 medium, showing a direct correlation between the media’s proline concentration and the in-vitro response. Interestingly, they also noticed that proline incorporation was notably higher with 15% FBS compared to 5 or 10% FBS.

Conclusions

The matrix cell density was found to be comparable between the 15% donor horse serum and the uncultured controls and demonstrated higher values than most other sera by the end of the second week. These findings suggest that the in-vitro culture system of the equine superficial digital flexor tendon (SDFT) tissue could be a useful tool in future studies aimed at boosting our understanding of the biological and biochemical characteristics associated with intrinsic tendon healing.