Journal: Nature Communications
Article Title: In vivo self-renewal and expansion of quiescent stem cells from a non-human primate
doi: 10.1038/s41467-025-58897-x
Figure Lengend Snippet: a BLI post-transplantation. Presumptive FAPs (NCAM1 − THY1 + ) were purified by FACS from three mouse lemurs, transduced with a lentivirus-expressing GFP and Luciferase, and transplanted into the TA muscles of NSG recipient mice. Mice were imaged over time and the normalized BLI signal was plotted as the mean plus standard error. n = 17 from 3 donors. b Freshly isolated FAPs (top panels) or MuSCs (bottom panels) from mice, lemur, or humans were transplanted into the TA muscles of NSG mice. One month post-transplantation, muscles were harvested, fixed, and frozen. Cryosections were stained with ORO. c Quantification of ORO stains shown in ( b ). n = 4 biological independent samples. The p -values were calculated by unpaired, two-tailed Student’s t -tests. *** p < 0.001 (p = 0.0005), * p < 0.05 ( p = 0.0445), ns = p > 0.05 (0.1573). d–f Freshly isolated FAPs were expanded in vitro and treated with differentiation media for adipogenic ( d ), fibrogenic ( e ), or osteogenic (f) conversion. Cells were stained for Perilipin-1 ( d ), SMA ( e ), or RUNX2 ( f ). Left panels show representative images, right panels show graphs of quantification. n = 3 biological independent samples. g Freshly isolated FAPs were stained for CFD protein levels. Left panels show representative images, right panel shows the graph of quantifications ( n = 3 biological independent samples). h Freshly isolated FAPs were transduced with luciferase-expressing lentivirus. Half of the cells were simultaneously treated with the CFD inhibitor Danicopan or vehicle (Ctrl). Cells were transplanted into the TA muscles of three recipient NSG mice. Engraftment was monitored by BLI. n = 6 from 2 donors. i Transplanted muscles from h were isolated, fixed, and stained for ORO. Left panels show representative images with arrowheads denoting ORO staining, right panels show graphs of quantification. n = 3 biological independent samples. All statistical data in this figure are presented as mean values ± SEM. The p -values in d – g were calculated by unpaired, one-tailed Student’s t -test and marked as *** p < 0.001, ** p < 0.01, * p < 0.05, ns = p > 0.05. The actual p -values were: d mouse vs. lemur p = 0.018, mouse vs. human p = 0.0247, lemur vs. human p = 0.1611; e mouse vs. lemur p = 0.0067, mouse vs. human p = 0.032, lemur vs. human p = 0.3781; f mouse vs. lemur p = 0.0003, mouse vs. human p = 0.0001, lemur vs. human p = 0.0006; g mouse vs. lemur p = 0.0111, mouse vs. human p = 0.0085, lemur vs. human p = 0.2536. The p -value in i was calculated by paired, two-tailed Student’s t -test (** p < 0.01, p = 0.0072). Source data are provided as a Source Data file.
Article Snippet: The CFD inhibitor Danicopan (MedChem Express, Cat no. HY-117930) was dissolved in DMSO (BP231, Fisher Scientific) and added to the adipogenic medium at a final concentration of 1 mM and replenished every 2 days for 2 weeks.
Techniques: Transplantation Assay, Purification, Transduction, Expressing, Luciferase, Muscles, Isolation, Staining, Two Tailed Test, In Vitro, One-tailed Test
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