Plating efficiency and grade of CD34 clones after culture and immediate recovery of single granulocytes
. | PB CD34 clone . | . | . | . | . | . | . | BM CD34 clone . | . | . | . | . | . | . | Single granulocyte mtDNA PCR . | . | . | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Donor (age, y/sex) . | Grade, no. . | . | . | . | . | . | . | Grade, no. . | . | . | . | . | . | . | . | PCR efficiency . | . | ||||||||||||||
| . | 1 . | 2 . | 3 . | 4 . | Subtotal, no. . | Microplates*/total wells . | PEf, % . | 1 . | 2 . | 3 . | 4 . | Subtotal, no. . | Microplates*/total wells . | PEf, % . | Granulocytes, no. . | No. positive . | % positive . | ||||||||||||||
| 1 (47/F) | 13 | 54 | 128 | 85 | 280 | 5/480 | 58.3 | 33 | 21 | 30 | 38 | 122 | 4/384 | 31.8 | 480 | 51 | 10.6 | ||||||||||||||
| 2 (38/F) | 33 | 61 | 64 | 34 | 192 | 4/384 | 50.0 | — | — | — | — | — | — | — | 1440 | 37 | 2.6 | ||||||||||||||
| 3 (43/M) | 24 | 20 | 23 | 26 | 93 | 3/288 | 32.3 | 279 | 149 | 91 | 127 | 646 | 20/1920 | 33.6 | 480 | 50 | 10.4 | ||||||||||||||
| 4 (34/M) | 38 | 38 | 58 | 66 | 200 | 5/480 | 41.7 | — | — | — | — | — | — | — | 768 | 111 | 14.5 | ||||||||||||||
| 5 (54/M) | 35 | 50 | 61 | 36 | 182 | 5/480 | 37.9 | 306 | 177 | 119 | 140 | 742 | 20/1920 | 38.6 | 768 | 42 | 5.5 | ||||||||||||||
| 6 (34/F) | 37 | 46 | 52 | 92 | 227 | 5/480 | 47.3 | — | — | — | — | — | — | — | 768 | 64 | 8.3 | ||||||||||||||
| Total | 180 | 269 | 386 | 339 | 1174 | 27/2592 | 45.3 | 618 | 347 | 240 | 305 | 1510 | 44/4224 | 35.7 | 4704 | 355 | 7.5 | ||||||||||||||
. | PB CD34 clone . | . | . | . | . | . | . | BM CD34 clone . | . | . | . | . | . | . | Single granulocyte mtDNA PCR . | . | . | ||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Donor (age, y/sex) . | Grade, no. . | . | . | . | . | . | . | Grade, no. . | . | . | . | . | . | . | . | PCR efficiency . | . | ||||||||||||||
| . | 1 . | 2 . | 3 . | 4 . | Subtotal, no. . | Microplates*/total wells . | PEf, % . | 1 . | 2 . | 3 . | 4 . | Subtotal, no. . | Microplates*/total wells . | PEf, % . | Granulocytes, no. . | No. positive . | % positive . | ||||||||||||||
| 1 (47/F) | 13 | 54 | 128 | 85 | 280 | 5/480 | 58.3 | 33 | 21 | 30 | 38 | 122 | 4/384 | 31.8 | 480 | 51 | 10.6 | ||||||||||||||
| 2 (38/F) | 33 | 61 | 64 | 34 | 192 | 4/384 | 50.0 | — | — | — | — | — | — | — | 1440 | 37 | 2.6 | ||||||||||||||
| 3 (43/M) | 24 | 20 | 23 | 26 | 93 | 3/288 | 32.3 | 279 | 149 | 91 | 127 | 646 | 20/1920 | 33.6 | 480 | 50 | 10.4 | ||||||||||||||
| 4 (34/M) | 38 | 38 | 58 | 66 | 200 | 5/480 | 41.7 | — | — | — | — | — | — | — | 768 | 111 | 14.5 | ||||||||||||||
| 5 (54/M) | 35 | 50 | 61 | 36 | 182 | 5/480 | 37.9 | 306 | 177 | 119 | 140 | 742 | 20/1920 | 38.6 | 768 | 42 | 5.5 | ||||||||||||||
| 6 (34/F) | 37 | 46 | 52 | 92 | 227 | 5/480 | 47.3 | — | — | — | — | — | — | — | 768 | 64 | 8.3 | ||||||||||||||
| Total | 180 | 269 | 386 | 339 | 1174 | 27/2592 | 45.3 | 618 | 347 | 240 | 305 | 1510 | 44/4224 | 35.7 | 4704 | 355 | 7.5 | ||||||||||||||
Numbers in columns indicate cells or clones observed microscopically after 5-day suspension culture of CD34 cells from PB and BM or immediate recovery of sorted granulocytes. PB indicates peripheral blood; BM, bone marrow; grade 1, less than 5 cells/well; grade 2, 6 to 10 cells/well; grade 3, 11 to 20 cells/well; grade 4, more than 21 cells/well; PEf, plating efficiency: number of wells with clones/total wells.
Number of 96-well microplates.