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/cs-CD146 and ~7.5% cs-DLK1/cs-CD34/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/CD146 cells. CD34/CD146 cells were slightly more bulky than CD34/CD146 cells. Both CD34/CD146 and CD34/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.

Enrichment of CD146+ Adipose-Derived Stem Cells in Combination with Articular Cartilage Extracellular Matrix Scaffold Promotes Cartilage Regeneration

“…Later, it was also recognized as a marker for pericytes 24, 26, which have been identified as the natural ancestors of MSCs 27, 28. In human tissues, CD146 + cell was regarded as the mural cells of blood micro vessels 29, 30. Previous reports also showed that CD146 was associated with cell migration and self-renewal 31, 32.…”

Evolution of ASC Immunophenotypical Subsets During Expansion In Vitro

“…Similarly, it is intriguing that a subset of these cells exhibiting the CD31−CD34+CD146− profile, previously found to identify a stem cell lineage [25,64,65], also grew proportionally to the culture period. In addition, possibly important ramifications may reveal that some antigens that have been ambiguously associated with stemness, including CD200, CD248, and CD271 [66,67], were reinforced during the culture, while all subsets featuring CD146, which has previously been recognized as a surrogate for stemness [40,[68][69][70][71] were sequestered. Concerning interpersonal variability, the spread of the data roughly follows previously reported trends [49,72], except for two specific subsets, the CD166+CD271−CD248+ and CD166+CD271+CD248+, which were identified only in a single donor.…”

CRISPR/Cas9-mediated gene knockout in human adipose stem/progenitor cells

“…ASCs were isolated and maintained as previously described [23]. Briefly, sWAT samples were handled under sterile conditions, washed twice with PBS and dissected.…”