Phenotypical and Functional Characterization of Freshly Isolated Adipose Tissue-Derived Stem Cells

Abstract

Adipose tissue contains a stromal vascular fraction (SVF) that is a rich source of adipose tissue-derived stem cells (ASCs). ASCs are multipotent and in vitro-expanded ASCs have the capacity to differentiate, into amongst others, adipocytes, chondrocytes, osteoblasts, and myocytes. For tissue engineering purposes, however, it would be advantageous to use the whole SVF, which can be transplanted without further in vitro selection or expansion steps. Because little is known about the freshly isolated ASCs in the SVF, we phenotypically characterized human freshly isolated ASCs, using flow cytometry. In addition, we investigated whether freshly isolated ASCs have functional properties comparable to cultured ASCs. For this, the differentiation potential of both freshly isolated ASCs and cultured ASCs into the osteogenic pathway was analyzed. Freshly isolated ASCs slightly differed in immunophenotype from cultured ASCs. Contrary to cultured ASCs, freshly isolated ASCs were shown to be highly positive for CD34, and positive for CD117 and HLA-DR. On the other hand, expression of CD105 and especially CD166 on the freshly isolated ASCs was relatively low. After osteogenic stimulation of freshly isolated ASCs, both Runx-2 and CollaI gene expression were significantly increased (p < 0.05). However, there was a difference in the kinetics of gene expression between freshly isolated and cultured ASCs and also between the different SVF isolates tested. There was no difference in alkaline phosphatase activity between freshly isolated ASCs and cultured ASCs. In addition, freshly isolated ASCs stained positive for osteonectin and showed matrix mineralization. We conclude that although there are minor differences in phenotype and kinetics of differentiation between freshly isolated ASCs and cultured ASCs, the use of freshly isolated ASCs for tissue engineering purposes involving bone repair is potentially applicable.

Stromal Vascular Fraction Cells of Adipose and Connective Tissue in People with Osteoarthritis: A Case Control Prospective Multi-Centric Non-Randomized Study

“…In addition, the adipose tissue contains unique populations of cells that suppress the inflammatory responses, and thus further contribute to tissue healing. Loose connective tissue creates an optimal environment for adaptation of stem cells and other cells contained in SVF that supports and positively affects microenvironment being sentinels for injury, damaged cells and tissues [10,28]. Adipose-derived stem and stromal cells do not require in vitro cultivation and are ready for use immediately after isolation from the adipose tissue.…”

Comparison of the Viability and Yield of Adipose-Derived Stem Cells (ASCs) from Different Donor Areas

“…In our study, we constantly compared cultured ASCs rather than freshly isolated ASCs considering their higher level of homogeneity. Due to the fact that cultured ASCs display minor differences in phenotype and kinetics of differentiation compared to freshly isolated ASCs (21) be hypothesized that both types of ASCs may exhibit the same viability profile.…”

Constant expression of hexose-6-phosphate dehydrogenase during differentiation of human adipose-derived mesenchymal stem cells

“…However, SVF represents a heterogeneous cell population (Varma et al 2007, De Sousa Peixoto et al 2008; therefore, we prepared mesenchymal precursor/stem cells from human adipose tissue ADMSCs, as these cells represent undifferentiated precursor of several mesenchymal lineages including chondrocytes, osteocytes, and adipocytes (Zuk et al 2001(Zuk et al , 2002. The differentiative potentials of these mesenchymal cells were confirmed following culture in specific media for differentiation in adipogenic, osteogenic, and chondrogenic phenotypes as detailed in the 'Materials and methods' section.…”