Table 2.
Summary of results from immunomagnetic cell selection
|
% CD135
|
FLT3 ratio
|
||||||
|---|---|---|---|---|---|---|---|
| Sample no. | Total unsorted cells | Total yield, CD34+/CD38− cells (% unsorted) | Unsorted cells | Sorted cells | Sorted, % CD34+/CD38− | Unsorted cells | Sorted cells |
| 1 | 109 | 4.5 × 106 (0.45) | 96.7 | 97.8 | 98.2 | 0.67 | 0.69 |
| 2 | 109 | 3 × 106 (0.30) | 95.2 | 96.5 | 74.6 | 0.70 | 0.73 |
| 3 | 8 × 108 | 1.5 × 106 (0.19) | 89.5 | 98.1 | 93.8 | 21.0 | ND |
| 4 | 4 × 108 | 1 × 106 (0.25) | 96.4 | 99.5 | 71.8 | 0.77 | 0.70 |
| 5 | 2 × 108 | 3.5 × 104 (0.02) | 97.7 | 98.9 | 73.7 | 0.67 | 0.79 |
| 6 | 2.5 × 108 | 20 × 106 (8.0) | 91.0 | 92.9 | 95.7 | 9.50 | ND |
| 7 | 2.5 × 108 | 2.4 × 106 (0.96) | 10.5 | 0.2 | 98 | 0.80 | 0.90 |
| 8 | 3 × 108 | 1.5 × 106 (0.5) | 5.2 | 0.4 | 99.1 | 0.65 | 0.77 |
AML samples were stained with anti-CD34 and anti-CD38 antibodies and subjected to immunomagnetic selection as described in “Materials and methods.” Total unsorted cells represents the starting sample size, whereas total yield CD34+/CD38− cells refers to the total number of CD34+/CD38− cells obtained from a given sample after selection. The cells were also stained with anti-CD135 to analyze for cell surface expression of FLT3. Genomic DNA from sorted and unsorted cells was isolated, PCR amplified, and analyzed for the FLT3 mutant-wild-type allelic ratio. ND indicates that no wild-type FLT3 was detected in the sample.