ZFIN Image: Paul et al., 2019, Fig. 6

Image

Figure Caption

Fig. 6

Primary human macrophages can survive at physiological temperature of the zebrafish in vivo and do not cause a sustained inflammatory response. (a) Representative light sheet micrographs of uninjected, control whole larva at 3 days post-fertilization (dpf). Both transgenic mpeg:GFP (macrophages-green)/flk:mCherry (vessels-red) (top) and mpx:GFP (neutrophils-green)/flk:mCherry (vessels-red) (bottom) are shown. (b) Representative lightsheet micrographs of head regions of uninjected, control larvae at 3 dpf. Both transgenic mpeg:GFP (macrophages-green)/flk:mCherry (vessels-red) (top) and mpx:GFP (neutrophils-green)/flk:mCherry (vessels-red) (bottom) are shown. (c) Schematic of experimental design: primary monocytes were differentiated into macrophages before injection into the zebrafish brain at age 2 dpf and imaged at 1 and 7 days post injection (dpi). (d) Plot (mean ± SD) of average in vivo survival calculated for primary human macrophages differentiated at physiological temperatures of the zebrafish or human obtained from 3 larvae each, where the numbers of cells that survived over the course of 7 days were normalized to the initial numbers measured one day post injection. Differences in survival were not significant by two-way ANOVA with Tukey’s multiple comparisons post-test. The temperature of macrophage differentiation (P = 0.3072), fish line injected (P = 0.4822), and interaction between factors (P = 0.4575) were not significant sources of variation by two-way ANOVA with one degree of freedom. Monocytes that had been cultured in the presence of human macrophage colony stimulating factor (H-M-CSF) for eight days (e) at physiological temperature of (28.5 °C) and (f) at physiological temperature of humans (37 °C). Primary human macrophages (blue) were injected into the hind brain of transgenic mpeg:GFP (macrophages-green)/flk:mCherry (vessels-red) or mpx:GFP (neutrophils-green)/flk:mCherry (vessels-red) zebrafish larvae at 2 dpf. Micrographs of 3D projections showing distribution and survival of human primary macrophages 1 dpi, when larvae are 3 dpf, and 7 dpi, when larvae are 9 dpf. The same fish is shown in the left and right panels.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Sci. Rep.