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1.
Figure 4

Figure 4. CD56 negative NK cell precursors are markedly enriched in UCB. From: Mouse fetal and embryonic liver cells differentiate human umbilical cord blood (UCB) progenitors into CD56 negative NK cell precursors in the absence of IL-15.

Fresh adult blood (n=10) and UCB (n=20) were depleted of CD3+ T cells and stained for CD56, CD3, CD34 and CD7. This representative phenotypic shows a 10-fold enrichment of CD56 negative NK precursors in UCB.

Valarie McCullar, et al. Exp Hematol. ;36(5):598-608.
2.
Figure 6

Figure 6. EL08-1D2 better supports NK cell precursor differentiation. From: Mouse fetal and embryonic liver cells differentiate human umbilical cord blood (UCB) progenitors into CD56 negative NK cell precursors in the absence of IL-15.

CD34+/Lin/CD38 cells were cultured on AFT024 (n=7) or EL08-1D2 (n=7) for 28 days. The importance of cell-cell contact was determined by culture in direct contact or separated from stroma by a Transwell (TW) membrane prohibiting cell-cell contact. CD56 negative NK cell precursor differentiation from 10 starting cells is reported after culture with IL-3 and Flt3L.

Valarie McCullar, et al. Exp Hematol. ;36(5):598-608.
3.
Figure 3

Figure 3. CD56 negative NK precursors give rise to CD56+ NK cells in secondary culture. From: Mouse fetal and embryonic liver cells differentiate human umbilical cord blood (UCB) progenitors into CD56 negative NK cell precursors in the absence of IL-15.

UCB CD34+/Lin/CD38 cells (n=5) were cultured for 28 days on AFT024 supplemented with IL-3 and FL and then sorted into CD34+/CD7, CD34+/CD7+, and CD34/CD7+ NK cell precursors and plated into a secondary culture with cytokines as indicated for an additional 28 days. The absolute cell number of CD56+ NK cells and their ability to express KIR and CD94 is shown.

Valarie McCullar, et al. Exp Hematol. ;36(5):598-608.
4.
Figure 1

Figure 1. NK cell development on AFT024, IL-3 and FL can occur in the absence of IL-15. From: Mouse fetal and embryonic liver cells differentiate human umbilical cord blood (UCB) progenitors into CD56 negative NK cell precursors in the absence of IL-15.

UCB CD34+/Lin/CD38 cells (n=10) were cultured at 50 cells/well in direct contact with irradiated AFT024 and the indicated cytokines. On day 28, cells were harvested and analyzed by flow cytometry for the number of CD56+ NK cells. Results are normalized to 10 starting cells.

Valarie McCullar, et al. Exp Hematol. ;36(5):598-608.
5.
Figure 2

Figure 2. CD56 negative NK cell precursors accumulate in the absence of IL-15. From: Mouse fetal and embryonic liver cells differentiate human umbilical cord blood (UCB) progenitors into CD56 negative NK cell precursors in the absence of IL-15.

UCB CD34+/Lin/CD38 cells (n=10) were cultured for 28 days on AFT024 and cytokines. A) Shown is a representative example of a culture supplemented with IL-3, IL-21 and Flt3L. Cultured progeny were gated on CD56 negative lymphocytes. The accumulation of (B) CD34+/CD7, (C) CD34+/CD7+, and (D) CD34/CD7+ precursors are shown for the indicated cytokines. Results are normalized to 10 starting cells.

Valarie McCullar, et al. Exp Hematol. ;36(5):598-608.
6.
Figure 5

Figure 5. EL08-1D2 is superior to AFT024 to support NK cell differentiation. From: Mouse fetal and embryonic liver cells differentiate human umbilical cord blood (UCB) progenitors into CD56 negative NK cell precursors in the absence of IL-15.

CD34+/Lin/CD38 cells were cultured on AFT024 (n=4) or EL08-1D2 (n=4) separated from stroma by a Transwell (TW) membrane prohibiting cell-cell contact. NK cell differentiation (measured by the CD56 expressing cells) from 10 starting cells was followed over time after culture with IL-15, IL-3, IL-7, KL, FL. For CD34+/Lin/CD38 cells in contact with EL08-1D2, counts are not shown beyond day 28 as cell numbers decreased. EL08-1D2 supported NK cell differentiation better than all other conditions tested (P=<0.03 for all points at days 21 and 28).

Valarie McCullar, et al. Exp Hematol. ;36(5):598-608.

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