The Adipose-Derived Lineage-Negative Cells Are Enriched Mesenchymal Stem Cells and Promote Limb Ischemia Recovery in Mice

Abstract

White adipose tissue (WAT) is a very attractive source of mesenchymal stem cells (MSCs) because of its availability and ease of harvest. However, the current method of isolating adipose tissue-derived MSCs often relies on the adhesiveness of the cultured stromal-vascular fraction (SVF). Unfortunately, the SVF is a heterogeneous cell population containing many cell types, including adipocyte precursor cells, endothelial cells, pericytes, multipotent MSCs, erythrocytes, and hematopoietic cells. Here we systematically characterized the adipose tissue-derived lineage-negative (Lin(-)) cell population using various surface markers and a set of cell proliferation and differentiation assays. We demonstrate clearly that the Lin(-) cell population represents enriched MSCs, which were identified by their high expression of MSC surface markers, and that these cells are a robust population with a vigorous growth capability and delayed aging. This cell population also demonstrated a much higher capacity for differentiation into osteogenic, chondrogenic and adipogenic cell lineages related to MSCs than did the SVF. These cells promoted recovery from limb ischemia, likely via production of vascular endothelial growth factor, an angiogenic factor. Our study demonstrates that Lin(-) cells are enriched in MSCs and provides a reliable method for isolating purer MSCs than SVF cells from the WAT, especially for obtaining fresh MSCs for clinical applications. In summary, this study identified a new, reliable method for enrichment of WAT MSCs with regenerative repairing features.

Generation of embryonic stem cells from mouse adipose-tissue derived cells via somatic cell nuclear transfer

“…18 Previously, we have used Lin ¡ cells to successfully generate cloned mice via SCNT. 17 We found that the rate of in vitro development of reconstructed oocytes into blastocysts was significantly higher from Lin ¡ cells than from Lin C cells.…”

Adipose tissue-derived therapeutic cells for peripheral artery diseases: the fatty blessing

“…In this view, selection of lineage-negative cells from adipose tissue has been shown to provide a reliable method for isolating higher number of more efficient ASCs than the use of SVF. [12] In addition, the source of fat could dictate human ASCs reparative activity. Hence, ASCs harvested from five different subcutaneous adipose depots in 12 female patients classified into three age ranges have been compared for their capacity of differentiation, proliferation, and sensitivity to apoptosis.…”

Conversion of Adipose Tissue-Derived Mesenchymal Stem Cells to Neural Stem Cell-Like Cells by a Single Transcription Factor, Sox2

“…In this study, we used our previously purified and characterized ADSC cell line, which expressed specific MSC markers and possessed osteogenic, chondrogenic, and adipogenic differentiation potential (Qin et al, 2014). When the ADSCs were cultured in normal medium (DMEM with 10% FBS), they exhibited a typical fibroblast-like morphology (Fig.…”

“…Adipose tissuederived mesenchymal stem cells (ADSCs) have multipotency, can undergo self-renewing divisions, and possess the capacity to differentiate into osteogenic, chondrogenic, and adipogenic cell lineages (Qin et al, 2014;Zeve et al, 2009;Zuk et al, 2001). In addition, human and mouse adipose tissuederived stem cells not only can be reprogrammed to induced pluripotent stem cells (iPSCs) with substantially higher efficiencies than those reported for human and mouse fibroblasts (Sugii et al, 2010), but they also have stronger proliferation and differentiation capabilities in vitro than skin fibroblasts (Rodeheffer et al, 2008;Zuk et al, 2001).…”

“…In this study, we used our previously purified and characterized ADSCs that expressed mesenchymal stem cell (MSC)-specific markers and possessed osteogenic, chondrogenic, and adipogenic differentiation potential (Qin et al, 2014). We found that the ADSCs maintained in monolayer or in spheres did not express Sox2 and Pax6 genes and proteins and did not differentiate to neuron-like cells.…”