Expansion of purified BM CD34+ subsets following coculture with HUBEC monolayers
. | . | . | Day 7 . | . | . | . | . | . | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | Day 0 . | . | Total . | . | CD34+ . | . | CD34+CD38- . | . | |||||
| Culture condition . | Cell subset . | Cell count, × 105 . | Cell count, × 105 . | Fold change . | Cell count, × 105 . | Fold change . | Cell count, × 105 . | Fold change . | |||||
| HUBECs (contact) + GM36SF | 34+ | 1.0 | 16.2 ± 0.3 | 16 | 3.9 ± 0.1 | 3.9 | 3.7 ± 0.1 | 115 | |||||
| HUBECs (contact) + GM36SF | 34+38+ | 1.0 | 18.3 ± 0.1 | 18 | 2.6 ± 0.1 | 2.6 | 2.4 ± 0.1 | 2.4 | |||||
| HUBECs (contact) + GM36SF | 34+38- | 0.4 | 2.7 ± 0.7 | 6.7 | 1.8 ± 0.5 | 4.5 | 1.8 ± 0.5 | 4.4 | |||||
| HUBECs (noncontact) + GM36SF | 34+38- | 0.4 | 6.7 ± 0.1 | 16.7 | 3.4 ± 0.5 | 8.6 | 2.5 ± 0.1 | 6.4 | |||||
| HUBECs (contact) + TSF | 34+38- | 0.4 | 5.8 ± 0.5 | 14.6 | 5.3 ± 0.4 | 13.4 | 5.2 ± 0.5 | 13.1 | |||||
| GM36SF | 34+38- | 0.4 | 14.4 ± 0.7 | 35.8 | 3.7 ± 0.2 | 9.2 | 0.3 ± 0.1 | — | |||||
| TSF | 34+38- | 0.4 | 1.4 ± 0.2 | 3.5 | 1.0 ± 0.1 | 2.4 | 0.1 ± 0.1 | — | |||||
. | . | . | Day 7 . | . | . | . | . | . | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| . | Day 0 . | . | Total . | . | CD34+ . | . | CD34+CD38- . | . | |||||
| Culture condition . | Cell subset . | Cell count, × 105 . | Cell count, × 105 . | Fold change . | Cell count, × 105 . | Fold change . | Cell count, × 105 . | Fold change . | |||||
| HUBECs (contact) + GM36SF | 34+ | 1.0 | 16.2 ± 0.3 | 16 | 3.9 ± 0.1 | 3.9 | 3.7 ± 0.1 | 115 | |||||
| HUBECs (contact) + GM36SF | 34+38+ | 1.0 | 18.3 ± 0.1 | 18 | 2.6 ± 0.1 | 2.6 | 2.4 ± 0.1 | 2.4 | |||||
| HUBECs (contact) + GM36SF | 34+38- | 0.4 | 2.7 ± 0.7 | 6.7 | 1.8 ± 0.5 | 4.5 | 1.8 ± 0.5 | 4.4 | |||||
| HUBECs (noncontact) + GM36SF | 34+38- | 0.4 | 6.7 ± 0.1 | 16.7 | 3.4 ± 0.5 | 8.6 | 2.5 ± 0.1 | 6.4 | |||||
| HUBECs (contact) + TSF | 34+38- | 0.4 | 5.8 ± 0.5 | 14.6 | 5.3 ± 0.4 | 13.4 | 5.2 ± 0.5 | 13.1 | |||||
| GM36SF | 34+38- | 0.4 | 14.4 ± 0.7 | 35.8 | 3.7 ± 0.2 | 9.2 | 0.3 ± 0.1 | — | |||||
| TSF | 34+38- | 0.4 | 1.4 ± 0.2 | 3.5 | 1.0 ± 0.1 | 2.4 | 0.1 ± 0.1 | — | |||||
Human BM CD34+ cells (99.1% ± 0.1% CD34+, 3.2% ± 0.1% CD34+CD38-, 96.0% ± 0.1% CD34+CD38+) were cultured with HUBECs supplemented with GM36SF (N = 3). FACS-sorted BM CD34+CD38- cells (N = 8) and BM CD34+CD38+ cells (N = 6) were placed in contact or noncontact cultures with HUBEC monolayers and cytokine supplements as indicated in “Materials and methods.” For comparison, BM CD34+CD38- cells were cultured in liquid suspension with either GM36SF or TSF (N = 3).
— indicates a decline in cells compared to input.