TABLE II.
Distribution of C2 Clones by Size and Staining with Antibodies against MyoD and Myogenina
| In 2% FCS |
In 2% FCS &
PDGF |
|||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 2 days |
3 days |
2 days |
3 days |
|||||||||
| Cloneb No. |
Clonec Size |
MyoD+d Cells |
Myogenin+ Cells |
Clone Size |
MyoD+ Cells |
Myogenin+ Cells |
Clone Size |
MyoD+ Cells |
Myogenin+ Cells |
Clone Size |
MyoD+ Cells |
Myogenin+ Cells |
| 1 | + | 0 | 0 | ++ | 0 | 0 | ++ | 0 | 0 | ++ | 0 | 0 |
| 2 | + | 0 | 0 | ++ | 0 | 0 | ++ | 0 | 0 | ++ | 0 | 0 |
| 3 | + | 0 | 0 | +++ | 0 | 0 | ++ | 0 | 0 | ++ | 1 | 0 |
| 4 | + | 0 | 0 | +++ | 0 | 0 | ++ | 0 | 0 | ++ | 3 | 1 |
| 5 | + | 0 | 0 | +++ | 0 | 0 | ++ | 0 | 0 | +++ | 0 | 0 |
| 6 | + | 0 | 0 | +++ | 1 | 0 | +++ | 0 | 0 | +++ | 0 | 0 |
| 7 | ++ | 0 | 0 | +++ | 2 | 1 | +++ | 0 | 0 | +++ | 0 | 0 |
| 8 | ++ | 0 | 0 | +++ | 3 | 3 | +++ | 0 | 0 | +++ | 0 | 0 |
| 9 | ++ | 0 | 0 | ++++ | 0 | 1 | +++ | 0 | 0 | +++ | 1 | 0 |
| 10 | ++ | 0 | 0 | ++++ | 1 | 1 | +++ | 0 | 0 | +++ | 3 | 0 |
| 11 | ++ | 0 | 0 | ++++ | 1 | 2 | +++ | 1 | 0 | +++ | 4 | 1 |
| 12 | +++ | 0 | 0 | ++++ | 1 | 2 | ++++ | 0 | 0 | ++++ | 0 | 0 |
| 13 | +++ | 0 | 0 | ++++ | 2 | 2 | ++++ | 0 | 0 | ++++ | 1 | 0 |
| 14 | +++ | 0 | 0 | ++++ | 2 | 4 | ++++ | 0 | 0 | ++++ | 1 | 1 |
| 15 | +++ | 0 | 0 | ++++ | 3 | 1 | ++++ | 0 | 0 | ++++ | 2 | 0 |
| 16 | +++ | 0 | 0 | ++++ | 3 | 1 | ++++ | 0 | 0 | ++++ | 2 | 0 |
| 17 | +++ | 0 | 0 | ++++ | 3 | 3 | ++++ | 0 | 0 | ++++ | 2 | 1 |
| 18 | +++ | 0 | 0 | ++++ | 3 | 3 | ++++ | 0 | 0 | ++++ | 2 | 1 |
| 19 | +++ | 0 | 0 | ++++ | 3 | 3 | ++++ | 0 | 0 | ++++ | 2 | 1 |
| 20 | ++++ | 0 | 0 | ++++ | 3 | 3 | ++++ | 0 | 0 | ++++ | 2 | 2 |
| 21 | ++++ | 1 | 0 | ++++ | 3 | 5 | ++++ | 1 | 0 | ++++ | 2 | 2 |
| 22 | ++++ | 1 | 0 | ++++ | 4 | 3 | ++++ | 1 | 0 | ++++ | 2 | 2 |
| 23 | ++++ | 1 | 1 | ++++ | 4 | 3 | ++++ | 2 | 1 | ++++ | 3 | 1 |
| 24 | ++++ | 2 | 1 | ++++ | 4 | 3 | ++++ | 2 | 1 | ++++ | 3 | 2 |
| 25 | ++++ | 3 | 1 | ++++ | 4 | 4 | ++++ | 2 | 1 | ++++ | 3 | 2 |
| 26 | ++++ | 3 | 2 | ++++ | 4 | 4 | ++++ | 3 | 1 | ++++ | 3 | 3 |
| 27 | ++++ | 4 | 2 | ++++ | 5 | 1 | ++++ | 3 | 1 | ++++ | 4 | 4 |
| 28 | ++++ | 4 | 2 | ++++ | 5 | 2 | ++++ | 3 | 3 | ++++ | 5 | 3 |
| 29 | ++++ | 5 | 3 | ++++ | 5 | 3 | ++++ | 4 | 3 | ++++ | 5 | 3 |
| 30 | ++++ | 5 | 3 | ++++ | 5 | 5 | ++++ | 5 | 1 | ++++ | 5 | 3 |
Clonal cultures were maintained for 5 days in proliferation medium containing 20% FCS and 0.5% chicken embryo extract, then switched into the serum-poor medium containing 2% FCS ± PDGF for 2 or 3 additional days.
Clones are arranged in the table by their size; within each size group clones are arranged first by the proportion of MyoD+ cells and second by the proportion of myogenin+ cells.
Clone size was determined based on examining the clone using a 16x objective. “+” represents clones which occupy less than 1 microscopic field. “++” represents clones which occupy about 1 microscopic field. “+++” represents clones ranging in size from more than 1 microscopic field up to 2 fields. “++++” represents clones which occupy more than 2 microscopic fields. Sparse peripheral cells were not included in this sizing method.
Numbers under MyoD+ cells and myogenin+ cells are based on visual estimates of positive cells within a clone using a 0 to 5 scale where 0 reflects no positives and 5 reflects over 80% positives.