LTC-IC Amplification in CD34+ CB Cells Cultured in LTC in the Presence of Various Cytokine Combinations
| Factors Added . | No. of Cells/100 Initial Nucleated Cells . | ||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | Nonadherent Cells . | CFU-C . | LTC-IC . | . | . | . | . | . | . | . | . | . | . | . | . | . | . | . | |||||||||||||||
| . | Start . | 2 wk . | 5 wk . | 10 wk . | 15 wk . | 20 wk . | Start . | 2 wk . | 5 wk . | 10 wk . | 15 wk . | 20 wk . | Start . | 2 wk . | 5 wk . | 10 wk . | 15 wk . | 20 wk . | . | . | . | . | . | . | . | . | . | . | . | . | . | . | . |
| Med | 100 | 380 | 0 | 0 | 0 | 0 | 13.2 | 0 | 0 | 0 | 0 | 0 | 0.6 | 0 | 0 | 0 | 0 | 0 | |||||||||||||||
| FL | 100 | 8,400 | 97,500 | 51,000 | 0 | 0 | 13.2 | 144 | 847 | 0 | 0 | 0 | 0.6 | 0.75 | 0.63 | 0 | 0 | 0 | |||||||||||||||
| TPO | 100 | 4,800 | 2,000 | 0 | 0 | 0 | 13.2 | 51.2 | 0 | 0 | 0 | 0 | 0.6 | 0 | 0 | 0 | 0 | 0 | |||||||||||||||
| FL + TPO | 100 | 9,400 | 197,000 | 1,025,000 | 89,700,000 | 1,074,790,000 | 13.2 | 206 | 2,947 | 154,091 | 2,873,468 | 21,936,205 | 0.6 | 96 | ND | 599 | 1,975 | 83,886 | |||||||||||||||
| KL + FL + TPO | 100 | 25,000 | 227,000 | 2,514,000 | 124,110,000 | 1,454,895,000 | 13.2 | 202 | 835 | 58,655 | 2,714,461 | 17,469,274 | 0.6 | 76.8 | ND | ND | ND | 31,457 | |||||||||||||||
| IL-3 + FL + TPO | 100 | 19,400 | 185,000 | 128,000 | 0 | 0 | 13.2 | 157 | 1,597 | 0 | 0 | 0 | 0.6 | 0.4 | 0 | 0 | 0 | 0 | |||||||||||||||
| Factors Added . | No. of Cells/100 Initial Nucleated Cells . | ||||||||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | Nonadherent Cells . | CFU-C . | LTC-IC . | . | . | . | . | . | . | . | . | . | . | . | . | . | . | . | |||||||||||||||
| . | Start . | 2 wk . | 5 wk . | 10 wk . | 15 wk . | 20 wk . | Start . | 2 wk . | 5 wk . | 10 wk . | 15 wk . | 20 wk . | Start . | 2 wk . | 5 wk . | 10 wk . | 15 wk . | 20 wk . | . | . | . | . | . | . | . | . | . | . | . | . | . | . | . |
| Med | 100 | 380 | 0 | 0 | 0 | 0 | 13.2 | 0 | 0 | 0 | 0 | 0 | 0.6 | 0 | 0 | 0 | 0 | 0 | |||||||||||||||
| FL | 100 | 8,400 | 97,500 | 51,000 | 0 | 0 | 13.2 | 144 | 847 | 0 | 0 | 0 | 0.6 | 0.75 | 0.63 | 0 | 0 | 0 | |||||||||||||||
| TPO | 100 | 4,800 | 2,000 | 0 | 0 | 0 | 13.2 | 51.2 | 0 | 0 | 0 | 0 | 0.6 | 0 | 0 | 0 | 0 | 0 | |||||||||||||||
| FL + TPO | 100 | 9,400 | 197,000 | 1,025,000 | 89,700,000 | 1,074,790,000 | 13.2 | 206 | 2,947 | 154,091 | 2,873,468 | 21,936,205 | 0.6 | 96 | ND | 599 | 1,975 | 83,886 | |||||||||||||||
| KL + FL + TPO | 100 | 25,000 | 227,000 | 2,514,000 | 124,110,000 | 1,454,895,000 | 13.2 | 202 | 835 | 58,655 | 2,714,461 | 17,469,274 | 0.6 | 76.8 | ND | ND | ND | 31,457 | |||||||||||||||
| IL-3 + FL + TPO | 100 | 19,400 | 185,000 | 128,000 | 0 | 0 | 13.2 | 157 | 1,597 | 0 | 0 | 0 | 0.6 | 0.4 | 0 | 0 | 0 | 0 | |||||||||||||||
Differential ability of FL, TPO, KL, and IL-3, alone or combined, to support different stages of hematopoiesis in long term stroma-free cultures suspension cultures of CD34+ CB cells. Cultures were initiated with 2 × 103 highly purified CD34+ CB cells and maintained as described in the Materials and Methods. Results show the mean of three to five experiments performed in duplicate after normalization of each result per 100 cells seeded into each type of culture. The column on the left shows the number of cells generated after progressively longer periods of time in liquid cultures. The central column shows the committed progenitor production in the same liquid cultures. The right-side column shows the generation and expansion of primitive stem cells (identified as LTC-IC).