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. 2021 May 8;44(4):560–566. doi: 10.1016/j.htct.2021.02.005

Figure 1.

Fig 1

Morphology and purity analysis of murine BM-MSCs cultured according to different protocols. (A) Cells cultured according to protocol 1 after 21 days of plating. Cells cultured with 10% FBS were rounder and smaller, compared with cells cultured with 20% FBS, which were more elongated, arranged in groups and presented more protrusions (arrows). These cells did not reach the necessary confluency to be analyzed by flow cytometry. (B) Cells cultured according to protocol 2 after 10 days of culture with 10% HS or 20% HS supplementation. Observe that the cultures containing 20% HS presented more elongated cells with a fibroblastic-like cell shape (arrows) and higher confluency, whereas the 10% HS condition generated fewer cells, of which most were round and dark indicating unviability (arrows). (C) The panel of cell surface markers consisted of antibodies anti-Ter119, anti-CD45, anti-CD31, anti-CD11b, anti-Sca1 and anti-CD44. The red histogram represents control cells and the blue histogram represents the cells incubated with the indicated antibodies. Cells cultured according to protocol 2 were positive for CD45, CD31, CD11b, Sca1 and CD44 and negative for Ter119. (D) Cells cultured under protocol 3 after 7 days in culture. Cells presented either an elongated shape (arrows) and were spread in the flask or exhibited a dark rounded shape organized in clusters. (E) Expression of the differentiation markers was similar to protocol 2. (F) Cells after 7 days of culture performed according to protocol 4, showing adherent cells with a slightly elongated shape (arrows) coexisting in the same flask with non-adherent cells (shadows of the cells floating in the media). (G) Cells cultured following this protocol were positive for all surface markers. (H) Cells cultured according to protocol 5 after 25 days of plating, showing a few small round cells spread in the flask and some spindle-shaped cells (arrows) organized in clusters and forming a monolayer (phase contrast image). (I) Similar to protocols 2 and 3, cells cultured according to protocol 5 were also positive for CD31, CD11b, Sca1 and CD44 and CD45 and negative for Ter119.