Isolation and culture of MSCs and stromal subsets from lipoaspirate. (A) SVF was obtained from human lipoaspirate specimens (n = 4 donors). An aliquot of SVF was directly seeded in tissue-culture plates for the isolation of conventional MSCs by plastic adherence. Another aliquot of SVF was processed for FACS sorting of DAPI⁻CD45⁻CD34⁻CD146⁺ perivascular cells and DAPI⁻CD45⁻CD34⁺CD146⁻ cells. (B) FACS analysis of cultured fat-derived MSCs, CD146⁺ perivascular cells, and CD146⁻ cells. After 9 passages in culture, MSCs retain a low percentage of CD146⁺ cells, while purified CD146⁺ perivascular cells and CD146⁻ cells retain a stable phenotype homogeneously positive and negative for CD146, respectively.