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. 2018 Mar 1;10(3):890–904. doi: 10.1016/j.stemcr.2018.01.011

Figure 2.

Figure 2

CD45−CD34+ CD31Lo Population Demonstrated Bipotential Characteristics

(A) Placenta-derived cells were cultured in different media for 2–3 weeks, DMEM supplemented with 10% fetal bovine serum (a standard mesenchymal stem cell [MSC] growth medium), endothelial growth medium (EGM2, a standard endothelial growth medium), skeletal and smooth muscle cell media, and MethoCult (a standard hematopoietic medium). None of CD45−CD34+ subpopulations could grow in smooth and skeletal muscle media, or displayed hematopoietic capacity. CD31Neg and CD31Lo cells gave rise to colonies in standard MSC growth medium.

(B) In vitro characterization of each population cultured in EGM2 using immunofluorescence at passage 2. Upper rows: CD31Neg-derived cells were negative for CD34, UEA-1, CD45, CD31, and CD144, and positive for α-SMA and vimentin. Middle rows: staining of the CD31Lo population resulted in heterogeneous cells expressing CD31, CD144, and UEA-1 in parts of the colony and α-SMA in other parts. Bottom rows: CD31Int-derived cells were negative for CD34, α-SMA, and CD45, but labeled with vimentin, UEA-1, CD31, and CD144. Scale bar, 20 μm.

(C) MSC derived from CD31Lo population on passage differentiated into adipocyte and osteoblast lineages after 2 weeks of culture in appropriate differentiation medium. Scale bar, 100 μm.

(D) Cell number in each subpopulation after culturing 10,000 fresh isolated and flow-sorted cells in EGM2 after 3 weeks (mean ± SD).