Immobilization on FN or RN and subsequent detachment does not affect the colony forming ability of CD34+cells
| Method of Attachment† . | Method of Release . | Colony Formation* . | |||
|---|---|---|---|---|---|
| Number of Colonies Per 2000 Cells . | Percent of Nonimmobilized Control . | ||||
| Erythroid . | Myeloid . | Erythroid . | Myeloid . | ||
| — | — | 70 ± 5 | 52 ± 6 | 100 ± 5 | 100 ± 6 |
| Con A | Pipetting | 75 ± 16 | 26 ± 2 | 108 ± 20 | 54 ± 4 |
| FN + TS2/16.2.1 mAb | Pipetting | 67 ± 4 | 43 ± 8 | 98 ± 6 | 84 ± 19 |
| RN | Peptide | 81 ± 18 | 53 ± 8 | 117 ± 23 | 102 ± 14 |
| Method of Attachment† . | Method of Release . | Colony Formation* . | |||
|---|---|---|---|---|---|
| Number of Colonies Per 2000 Cells . | Percent of Nonimmobilized Control . | ||||
| Erythroid . | Myeloid . | Erythroid . | Myeloid . | ||
| — | — | 70 ± 5 | 52 ± 6 | 100 ± 5 | 100 ± 6 |
| Con A | Pipetting | 75 ± 16 | 26 ± 2 | 108 ± 20 | 54 ± 4 |
| FN + TS2/16.2.1 mAb | Pipetting | 67 ± 4 | 43 ± 8 | 98 ± 6 | 84 ± 19 |
| RN | Peptide | 81 ± 18 | 53 ± 8 | 117 ± 23 | 102 ± 14 |
Colonies formed were evaluated and designated as erythroid- or myeloid-containing. Colony formation is represented as number of colonies formed per 2000 cells plated and percentage of the nonimmobilized control (number of colonies formed by treated cells per number of colonies formed by control suspension cells × 100) ± SD for 4 separate experiments.
CD34 cells were incubated in Stem Cell Media 48 hours before onset of assay and then attached to matrix coated plates for 45 minutes. Cells were detached, resuspended in MethoCult GF Kit (StemCell Technologies, Vancouver, BC, Canada) (1000 cells/mL/plate), and incubated for 14 days in a humidified incubator at 37°C, 6% CO2.