CD146 (MCAM) in human cs-DLK1-/cs-CD34+ adipose stromal/progenitor cells

Abstract

To precisely characterize CD146 in adipose stromal/progenitor cells (ASCs) we sorted the stromal vascular faction (SVF) of human abdominal subcutaneous white adipose tissue (sWAT) according to cell surface (cs) expression of CD146, DLK1 and CD34. This test identified three main SVF cell populations: ~50% cs-DLK1^-/cs-CD34⁺/cs-CD146^- ASCs, ~7.5% cs-DLK1⁺/cs-CD34^dim/+/cs-CD146⁺ and ~7.5% cs-DLK1⁺/cs-CD34^dim/+/cs-CD146^- cells. All cells contained intracellular CD146. Whole mount fluorescent IHC staining of small vessels detected CD146⁺ endothelial cells (CD31⁺/CD34⁺/CD146⁺) and pericytes (CD31^-/CD34^-/CD146⁺ ASCs). The cells in the outer adventitial layer showed the typical ASC morphology, were strongly CD34⁺ and contained low amounts of intracellular CD146 protein (CD31^-/CD34⁺/CD146⁺). Additionally, we detected wavy CD34^-/CD146⁺ and CD34^dim/CD146⁺ cells. CD34^dim/CD146⁺ cells were slightly more bulky than CD34^-/CD146⁺ cells. Both CD34^-/CD146⁺ and CD34^dim/CD146⁺ cells were detached from the inner pericyte layer and protruded into the outer adventitial layer. Cultured early passage ASCs contained low levels of CD146 mRNA, which was expressed in two different splicing variants, at a relatively high amount of the CD146-long form and at a relatively low amount of the CD146-short form. ASCs contained low levels of CD146 protein, which consisted predominantly long form and a small amount of short form. The CD146 protein was highly stable, and the majority of the protein was localized in the Golgi apparatus. In conclusion, the present study contributes to a better understanding of the spatial localization of CD34⁺/CD146⁺ and CD34^-/CD146⁺ cells in the adipose niche of sWAT and identifies CD146 as intracellular protein in cs-DLK1^-/cs-CD34⁺/cs-CD146^- ASCs.

Keywords: Adipose stromal/progenitor cells; CD146; CD31; CD34; DLK1 (Pref1); Subcellular localization; Turnover.