Effect of Withdrawal of TGF-β1 on the Number and Viability mBMMC and on the Expression and Release of mMCP-1
| . | Cells (×105/mL) . | % Viability . | % Mast Cells . | % mMCP-1+ . | mMCP-1 (ng/mL) . | mMCP-1 (ng/106 cells) . |
|---|---|---|---|---|---|---|
| Day 2 | ||||||
| T/I/W/S | 16.5 ± 0.5 | 88 ± 0.9 | 97 ± 1.8 | 96 ± 4.0 | 1,849 ± 351* | 2,155 ± 305 |
| I/W/S | 18.1 ± 1.4 | 81 ± 1.7† | 99 ± 0.7 | 89 ± 3.0 | 59 ± 4.0‡ | 565 ± 65‡ |
| W/S | 22.8 ± 1.9† | 86 ± 1.0 | 99 ± 0.4 | 90 ± 2.6 | 100 ± 47‡ | 464 ± 65‡ |
| T/W/S | 17.8 ± 2.4 | 85 ± 2.7 | 98 ± 0.9 | 95 ± 0.5 | 977 ± 46‡ | 2,715 ± 725 |
| T/I/S | 13.9 ± 3.0 | 79 ± 1.1† | 98 ± 0.5 | 98 ± 0.8 | 1,476 ± 109 | 5,061 ± 660‡ |
| Day 4 | ||||||
| T/I/W/S | 8 ± 0.8 | 90 ± 1.9 | 99 ± 0.2 | 99 ± 0.5 | 1,935 ± 376* | 6,038 ± 466 |
| I/W/S | 6.5 ± 0.6 | 87 ± 3.4 | 99 ± 0.7 | 60 ± 1.3† | 75 ± 23† | 167 ± 24† |
| W/S | 6.7 ± 0.2 | 83 ± 2.0 | 99 ± 0.3 | 55 ± 28† | 90 ± 7† | 140 ± 11† |
| T/W/S | 5.3 ± 0.8 | 80 ± 4.1 | 98 ± 0.7 | 93 ± 1.4† | 908 ± 97† | 3,365 ± 330† |
| T/I/S | 4.1 ± 0.4† | 66 ± 3.3† | 99 ± 0.3 | 98 ± 0.2 | 2,460 ± 301 | 10,320 ± 1,216† |
| . | Cells (×105/mL) . | % Viability . | % Mast Cells . | % mMCP-1+ . | mMCP-1 (ng/mL) . | mMCP-1 (ng/106 cells) . |
|---|---|---|---|---|---|---|
| Day 2 | ||||||
| T/I/W/S | 16.5 ± 0.5 | 88 ± 0.9 | 97 ± 1.8 | 96 ± 4.0 | 1,849 ± 351* | 2,155 ± 305 |
| I/W/S | 18.1 ± 1.4 | 81 ± 1.7† | 99 ± 0.7 | 89 ± 3.0 | 59 ± 4.0‡ | 565 ± 65‡ |
| W/S | 22.8 ± 1.9† | 86 ± 1.0 | 99 ± 0.4 | 90 ± 2.6 | 100 ± 47‡ | 464 ± 65‡ |
| T/W/S | 17.8 ± 2.4 | 85 ± 2.7 | 98 ± 0.9 | 95 ± 0.5 | 977 ± 46‡ | 2,715 ± 725 |
| T/I/S | 13.9 ± 3.0 | 79 ± 1.1† | 98 ± 0.5 | 98 ± 0.8 | 1,476 ± 109 | 5,061 ± 660‡ |
| Day 4 | ||||||
| T/I/W/S | 8 ± 0.8 | 90 ± 1.9 | 99 ± 0.2 | 99 ± 0.5 | 1,935 ± 376* | 6,038 ± 466 |
| I/W/S | 6.5 ± 0.6 | 87 ± 3.4 | 99 ± 0.7 | 60 ± 1.3† | 75 ± 23† | 167 ± 24† |
| W/S | 6.7 ± 0.2 | 83 ± 2.0 | 99 ± 0.3 | 55 ± 28† | 90 ± 7† | 140 ± 11† |
| T/W/S | 5.3 ± 0.8 | 80 ± 4.1 | 98 ± 0.7 | 93 ± 1.4† | 908 ± 97† | 3,365 ± 330† |
| T/I/S | 4.1 ± 0.4† | 66 ± 3.3† | 99 ± 0.3 | 98 ± 0.2 | 2,460 ± 301 | 10,320 ± 1,216† |
Data are expressed as the mean ± SE (n = 4, unless otherwise stated). Data significantly different from the results for control cultures grown in the presence of T/I/W/S are indicated. mBMMC were grown in the presence of WEHI (15%)/rmIL-9 (5 ng/mL)/rrSCF (50 ng/mL) and rhTGF-β1 (1 ng/mL) for 7 days. These cultures yielded 15.2 × 105 mBMMC/mL, 88% viable, and 99.6% mMCP-1+, with 547 ng mMCP-1/mL of supernatant. Cells were transferred in quadruplicate at 5 × 105/mL to 48-well plates and cultured in varying combinations of cytokines as shown: T, rhTGF-β1 (1 ng/mL); I, rmIL-9 (5 ng/mL); W, WEHI-3B IL-3–rich supernatant (15%); S, rrSCF (50 ng/mL). The table shows the following parameters that were measured 2 and 4 days after transfer into cytokines: cell numbers and viability, percentage of mast cells in Leishman’s stained cytosmears, percentage of mMCP-1+ mast cells, and concentrations of mMCP-1 in culture supernatants (in nanograms per milliliter) and cell pellets (in nanograms per 106 cells) assessed by ELISA.
n = 3.
P < .03 (Mann Whitney).
P < .05 (Mann Whitney).