Comparison of the frequency of SRCs by limiting dilution assay
Route . | No. cells injected . | No. engrafted mice . | No. transplantations . |
---|---|---|---|
iBM | 10 000 | 2 | 2 |
1 000 | 6 | 6 | |
500 | 4 | 4 | |
250 | 5 | 5 | |
100 | 8 | 10 | |
50 | 7 | 12 | |
10 | 5 | 10 | |
5 | 2 | 9 | |
Intravenous | 10 000 | 2 | 2 |
5 000 | 5 | 5 | |
2 000 | 3 | 4 | |
500 | 3 | 5 | |
100 | 2 | 5 | |
50 | 1 | 5 |
Route . | No. cells injected . | No. engrafted mice . | No. transplantations . |
---|---|---|---|
iBM | 10 000 | 2 | 2 |
1 000 | 6 | 6 | |
500 | 4 | 4 | |
250 | 5 | 5 | |
100 | 8 | 10 | |
50 | 7 | 12 | |
10 | 5 | 10 | |
5 | 2 | 9 | |
Intravenous | 10 000 | 2 | 2 |
5 000 | 5 | 5 | |
2 000 | 3 | 4 | |
500 | 3 | 5 | |
100 | 2 | 5 | |
50 | 1 | 5 |
NOD/SCID mice (n = 84) were transplanted with various numbers of Lin−/low CD34+CD38− CB cells by iBM or intravenous injection. Engraftment in murine BM was analyzed at 8 weeks. Mice were considered to be engrafted when they were determined positive by both FACS and PCR analysis. The frequency of engrafting cells in CB Lin−/lowCD34+CD38− cells transplanted by iBM and intravenously was found to be 1 in 44 Lin−/lowCD34+CD38− cells (95% confidence limits, 1 in 27 to 1 in 70) and 1 in 660 Lin−/lowCD34+CD38− cells (95% confidence limits, 1 in 289 to 1 in 1510), respectively. SRC frequencies were calculated using Poisson statistics and the method of maximum likelihood with the assistance of the L-Calc software (StemCell Technologies).