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. Author manuscript; available in PMC: 2020 Dec 22.
Published in final edited form as: Stem Cells. 2019 Dec 20;38(4):530–541. doi: 10.1002/stem.3133

Figure 1: Donor SMA9 cells express mesenchymal progenitor markers and can differentiate into Col2.3GFP+ osteoblasts.

Figure 1:

A) SMA9 mice, labeling BMSCs, were crossed with Col2.3GFP mice, labeling mature osteoblasts and osteocytes, to generate SMA9/Col2.3GFP mice. n=3 male for all of Figure 1.

B) Bone marrow stromal cells from these mice were cultured, and after Day 7, osteogenic differentiation was induced. Col2.3GFP+ cells are largely absent at Day 7, begin to be present at Day 14 with differentiation, and are strongly present at Day 21. Note the co-localization of Col2.3GFP cells to mineralized regions in the brightfield (BF) images. Merged images at day 14 and day 21 contain dual positive yellow cells, indicating that αSMA labeled progenitors differentiated into Col2.3GFP expressing osteoblasts. αSMA+ cells were labeled by treating the culture with 4-OH tamoxifen on Days 4 and 6.

C) As Day 7 cells were used for transplantation into OIM mice, these cells were profiled for progenitor markers by flow cytometry. Within the (CD45/CD31/Ter119)− SMA9+ population, approximately 37.8% of cells were dual positive for mesenchymal and osteoprogenitor markers Sca-1 and CD51, while majority of cells were CD51+ (>90%). Representative dot plot is shown.