Fig. 10.
Fig. 10. Long-term incubation with NGF enhances chymase immunoreactivity in human umbilical cord blood-derived mast cells. Human umbilical cord blood-derived mast cells (19 weeks after start of culture; < 99% purity by tryptase immunostaining) were maintained for an additional 3 weeks in culture with or without purified NGF at 500 ng/mL (see text). At the start of culture, tryptase immunoreactivity (red reaction product) is detectable in essentially all of the mast cells (A, B). (B) is a higher magnification view of some of the same cells shown in (A); the arrows in (A and B) depict the same two cells, both of which appear to have bilobed nuclei (a common finding in such in vitro-derived mast cells). (C and D) Mast cells that were maintained for an additional 3 weeks without NGF exhibit a range of chymase immunoreactivity (red reaction product), from none (eg, “-” in D), to 1 + or 2 + in intensity (examples depicted in D). (D) is a higher magnification view of some of the same cells shown in (C), some of which have bilobed (small arrow) or multilobed (thick arrow) configurations. (E and F ) Mast cells that were maintained for an additional 3 weeks with NGF exhibit increased chymase immunoreactivity (red reaction product), with some cells of 3 + staining intensity, as well as those of 1 + or 2 + intensity (examples indicated in F ). (F ) is a higher magnification view of some of the same cells as in (E), some of which have bilobed (small arrow) or multilobed (thick arrow) nuclear configurations. (G and H) Negative control (a mouse monoclonal IgG1K antibody of irrelevant antigen specificity, rather than either the mouse monoclonal IgG1K antitryptase or antichymase antibody, was used as the first antibody) showing lack of nonspecific immunoreactivity in these mast cells (these were aliquots of the same cell population depicted in C and D). (H) is a higher magnification view of some of the same cells shown in (G), some of which have multilobed nuclei (thick arrows), a common finding in such in vitro-derived mast cells. These cells are from Experiment 1 in Fig 9. Scale bars in G (for A, C, E and G) and H (for B, D, F, and H) represent 10 μm.

Long-term incubation with NGF enhances chymase immunoreactivity in human umbilical cord blood-derived mast cells. Human umbilical cord blood-derived mast cells (19 weeks after start of culture; < 99% purity by tryptase immunostaining) were maintained for an additional 3 weeks in culture with or without purified NGF at 500 ng/mL (see text). At the start of culture, tryptase immunoreactivity (red reaction product) is detectable in essentially all of the mast cells (A, B). (B) is a higher magnification view of some of the same cells shown in (A); the arrows in (A and B) depict the same two cells, both of which appear to have bilobed nuclei (a common finding in such in vitro-derived mast cells). (C and D) Mast cells that were maintained for an additional 3 weeks without NGF exhibit a range of chymase immunoreactivity (red reaction product), from none (eg, “-” in D), to 1 + or 2 + in intensity (examples depicted in D). (D) is a higher magnification view of some of the same cells shown in (C), some of which have bilobed (small arrow) or multilobed (thick arrow) configurations. (E and F ) Mast cells that were maintained for an additional 3 weeks with NGF exhibit increased chymase immunoreactivity (red reaction product), with some cells of 3 + staining intensity, as well as those of 1 + or 2 + intensity (examples indicated in F ). (F ) is a higher magnification view of some of the same cells as in (E), some of which have bilobed (small arrow) or multilobed (thick arrow) nuclear configurations. (G and H) Negative control (a mouse monoclonal IgG1K antibody of irrelevant antigen specificity, rather than either the mouse monoclonal IgG1K antitryptase or antichymase antibody, was used as the first antibody) showing lack of nonspecific immunoreactivity in these mast cells (these were aliquots of the same cell population depicted in C and D). (H) is a higher magnification view of some of the same cells shown in (G), some of which have multilobed nuclei (thick arrows), a common finding in such in vitro-derived mast cells. These cells are from Experiment 1 in Fig 9. Scale bars in G (for A, C, E and G) and H (for B, D, F, and H) represent 10 μm.

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