Progenitor Content of Lin−CD34−CD38−Populations
| Assay Conditions . | Sample . | No.of Sorts* . | Cells/ Dish . | Colonies/ MC Dish . | CFC/103 Lin−CD34−CD38− Cells . |
|---|---|---|---|---|---|
| Serum-containing MC (standard CFC) assays† | mPB 1 mPB 2 BM 1 BM 2 BM 3 BM 4 BM 8 | 1 1 1 1 1 2 2 | 500 5,000 500 1,000 500 5,000 1,000 | 0, 0 3, 5 0, 0 0, 0 0, 0 6, 0 0, 0 | <1 <1 <1 <1 <1 <1 <1 |
| Serum-free MC assays‡ | BM 2 BM 3 | 1 1 | 1,000 500 | 0, 0 0, 0 | <1 <1 |
| Serum-containing MC assays of cells from 6-wk-old LTC2-153 | mPB 1 mPB 2 BM 1 BM 2 BM 3 BM 4 BM 8 | 1 1 1 1 1 2 2 | 500 5,000 500 1,000 500 5,000 1,000 | 0, 0 0, 0 0, 0 0, 0 0, 0 90, 68 0, 0 | <1 <0.1 <1 <0.5 <1 160 <0.5 |
| Assay Conditions . | Sample . | No.of Sorts* . | Cells/ Dish . | Colonies/ MC Dish . | CFC/103 Lin−CD34−CD38− Cells . |
|---|---|---|---|---|---|
| Serum-containing MC (standard CFC) assays† | mPB 1 mPB 2 BM 1 BM 2 BM 3 BM 4 BM 8 | 1 1 1 1 1 2 2 | 500 5,000 500 1,000 500 5,000 1,000 | 0, 0 3, 5 0, 0 0, 0 0, 0 6, 0 0, 0 | <1 <1 <1 <1 <1 <1 <1 |
| Serum-free MC assays‡ | BM 2 BM 3 | 1 1 | 1,000 500 | 0, 0 0, 0 | <1 <1 |
| Serum-containing MC assays of cells from 6-wk-old LTC2-153 | mPB 1 mPB 2 BM 1 BM 2 BM 3 BM 4 BM 8 | 1 1 1 1 1 2 2 | 500 5,000 500 1,000 500 5,000 1,000 | 0, 0 0, 0 0, 0 0, 0 0, 0 90, 68 0, 0 | <1 <0.1 <1 <0.5 <1 160 <0.5 |
Abbreviations: mPB, mobilized peripheral blood; BM, bone marrow (numbers specify sample numbers).
No. of sorts distinguishes experiments in which cells were plated directly after being sorted once (1) or after the lin−CD34−CD38− fraction collected from the first sort was resorted (2) before culture in methylcellulose (MC) or LTC.
Cells plated in 2 replicate methylcellulose cultures containing 30% FCS and 50 ng/mL SF; 20 ng/mL GM-CSF, G-CSF, IL-3, and IL-6; and 3 U/mL Epo as described in Materials and Methods.
These methylcellulose cultures contained a serum substitute instead of FCS and 100 ng/mL FL, 100 ng/mL SF, 50 ng/mL TPO, 20 ng/mL IL-3, and 10 ng/mL H–IL-6. They were thus identical in composition to the medium used for the 10-day liquid suspension cultures described in Materials and Methods except for the addition of an equivalent concentration of methylcellulose to that used in the standard CFC assays.
Cells/dish shows the number of cells placed into each LTC-IC assay culture. Six weeks later, the entire LTC was harvested and 10% of the cells assayed per CFC assay dish. Colonies/dish indicates the number of colonies seen in each of 2 replicate CFC assays.