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. 2023 Jan 9;10:1094124. doi: 10.3389/fcell.2022.1094124

FIGURE 4.

FIGURE 4

Immunocytochemical labelling confirms CD146+ cells are a rare subpopulation when expanded in vitro, and can be enriched via MACS. Tendon cell-surface markers CD44 (A), CD90 (B) and CD146 (C) demonstrate that CD146 cells are a rare subpopulation amongst cultured TDCs (passage 1) when compared to cells expressing CD44 or CD90. n = 3 (biological replicates). Scale bar = 100 µm. Immunocytochemically labelled CD146 cells (D; passage 2) labelled with CD146 reaffirmed that expression persisted once expanded in vitro. n = 3 (biological replicates). Scale bar = 100 µm. Flow cytometry (E,F) confirmed CD146 expression in CD146+ cell populations. Total events = 10,000. (G) The percentage of CD146 cells (approx. 2%) yields by MACS from the original tendon-derived cell suspensions as determined by cell counting, reiterated the rarity of CD146 cells. n = 9 (biological replicates). Across MACS isolations (H), flow cytometry confirms CD146 enrichment, albeit approximately 15% of cells in the CD146− negative fraction were positive for CD146. n = 2 (biological replicates) per cell fraction. Graphs were plotted as mean (µ) ± SD.