Effect of Pre-Tp Serum and Neutralization of Cytokines on MK Ploidy Distribution
| Serum Added . | Percentage of MK in Each Ploidy Class . | |||||
|---|---|---|---|---|---|---|
| . | 2 N . | 4 N . | 8 N . | 16 N . | 32 N . | 64 N . |
| Normal mare | 3.56 ± 6.2 | 14.21 ± 3.5 | 24.66 ± 5.8 | 20.48 ± 4.9 | 10.58 ± 4.4 | 2.97 ± 0.9 |
| Preinfection | 3.83 ± 2.5 | 15.11 ± 5.8 | 23.81 ± 9.2 | 19.24 ± 6.0 | 11.53 ± 5.6 | 2.38 ± 1.2 |
| Preinfection + anti–TGF-β | 4.97 ± 4.7 | 13.89 ± 5.5 | 25.03 ± 7.1 | 18.68 ± 4.7 | 12.24 ± 5.3 | 3.01 ± 0.8 |
| Preinfection + anti–TNF-α | 3.75 ± 5.2 | 15.28 ± 3.7 | 23.52 ± 6.6 | 20.34 ± 5.2 | 10.75 ± 4.9 | 2.34 ± 1.7 |
| Preinfection + anti–TGF-β and anti–TNF-α | 4.02 ± 3.6 | 14.25 ± 4.3 | 24.66 ± 5.8 | 22.56 ± 5.5 | 11.89 ± 4.5 | 2.74 ± 1.9 |
| Pre-Tp | 3.14 ± 2.0 | 14.39 ± 5.4 | 22.91 ± 9.4 | 18.98 ± 6.7 | 11.03 ± 5.7 | 2.78 ± 2.1 |
| Pre-Tp + anti–TGF-β | 4.30 ± 2.8 | 15.77 ± 6.1 | 24.86 ± 8.2 | 19.81 ± 5.7 | 10.64 ± 4.1 | 2.09 ± 1.5 |
| Pre-Tp + anti–TNF-α | 4.76 ± 1.0 | 14.14 ± 2.7 | 23.97 ± 3.6 | 24.10 ± 5.0 | 10.69 ± 4.1 | 2.03 ± 0.6 |
| Pre-Tp + anti–TGF-β and TNF-α | 5.01 ± 3.1 | 15.04 ± 5.6 | 23.09 ± 7.8 | 21.55 ± 5.2 | 12.47 ± 4.8 | 2.41 ± 1.1 |
| Pre-Tp + antibody control | 3.76 ± 6.1 | 15.38 ± 5.9 | 22.79 ± 6.5 | 23.69 ± 5.8 | 12.58 ± 5.3 | 2.66 ± 2.0 |
| Serum Added . | Percentage of MK in Each Ploidy Class . | |||||
|---|---|---|---|---|---|---|
| . | 2 N . | 4 N . | 8 N . | 16 N . | 32 N . | 64 N . |
| Normal mare | 3.56 ± 6.2 | 14.21 ± 3.5 | 24.66 ± 5.8 | 20.48 ± 4.9 | 10.58 ± 4.4 | 2.97 ± 0.9 |
| Preinfection | 3.83 ± 2.5 | 15.11 ± 5.8 | 23.81 ± 9.2 | 19.24 ± 6.0 | 11.53 ± 5.6 | 2.38 ± 1.2 |
| Preinfection + anti–TGF-β | 4.97 ± 4.7 | 13.89 ± 5.5 | 25.03 ± 7.1 | 18.68 ± 4.7 | 12.24 ± 5.3 | 3.01 ± 0.8 |
| Preinfection + anti–TNF-α | 3.75 ± 5.2 | 15.28 ± 3.7 | 23.52 ± 6.6 | 20.34 ± 5.2 | 10.75 ± 4.9 | 2.34 ± 1.7 |
| Preinfection + anti–TGF-β and anti–TNF-α | 4.02 ± 3.6 | 14.25 ± 4.3 | 24.66 ± 5.8 | 22.56 ± 5.5 | 11.89 ± 4.5 | 2.74 ± 1.9 |
| Pre-Tp | 3.14 ± 2.0 | 14.39 ± 5.4 | 22.91 ± 9.4 | 18.98 ± 6.7 | 11.03 ± 5.7 | 2.78 ± 2.1 |
| Pre-Tp + anti–TGF-β | 4.30 ± 2.8 | 15.77 ± 6.1 | 24.86 ± 8.2 | 19.81 ± 5.7 | 10.64 ± 4.1 | 2.09 ± 1.5 |
| Pre-Tp + anti–TNF-α | 4.76 ± 1.0 | 14.14 ± 2.7 | 23.97 ± 3.6 | 24.10 ± 5.0 | 10.69 ± 4.1 | 2.03 ± 0.6 |
| Pre-Tp + anti–TGF-β and TNF-α | 5.01 ± 3.1 | 15.04 ± 5.6 | 23.09 ± 7.8 | 21.55 ± 5.2 | 12.47 ± 4.8 | 2.41 ± 1.1 |
| Pre-Tp + antibody control | 3.76 ± 6.1 | 15.38 ± 5.9 | 22.79 ± 6.5 | 23.69 ± 5.8 | 12.58 ± 5.3 | 2.66 ± 2.0 |
Equine marrow cells were grown in liquid culture and prepared for ploidy analysis on day 10. Cells were labeled with a murine anti-GPIIbIIIa monoclonal antibody and propidium iodide and analyzed by flow cytometry. Between 2,000 and 3,000 MK were analyzed. Data shown are means from 16 foals, with triplicate cultures for each treatment. There were no significant ploidy distribution differences between cultures grown in the presence of preinfection or Pre-Tp serum. The antibody control category is pooled data from cultures receiving both antibody controls (murine isotype-matched monoclonal antibody and rabbit polyclonal antiserum) separately and in combination.