Isolation of Adipose-Derived Stromal Cells Without Enzymatic Treatment: Expansion, Phenotypical, and Functional Characterization

Abstract

Stem cell therapy is a potential method for the treatment of numerous diseases. The most frequent cellular source is bone-marrow-derived mesenchymal stromal cells (BM-MSCs). Human adipose-derived stromal cells (ADSCs) share similar properties with BM-MSCs as they support hematopoiesis, modulate ongoing immune responses, and differentiate into cells of mesodermal origin. On the other hand, ADSCs have higher frequency in situ, higher availability, and very few ethical issues compared with BM-MSCs, giving them an advantage over BM-MSCs for clinical use. Most of the methods used to isolate ADSCs contain a collagenase digestion step, but the type of collagenase and time of sample digestion vary among studies and these differences could have an impact on the cell properties and thus in result comparison. To overcome this obstacle, we propose a new method to isolate ADSCs from lipoaspirate without collagenase digestion step. We compared ADSCs obtained with our method versus classical protocol using collagenase digestion. Cells obtained with our method are equivalent but they have a better long-term hematopoietic support than those obtained with classical method. Moreover, our method has an advantage over the classical one as it is easier, safer, faster, less expensive, and more consistent with good manufacturing practices to obtain large number of ADSCs ex vivo.

Characterization of Chondrogenic Gene Expression and Cartilage Phenotype Differentiation in Human Breast Adipose-Derived Stem Cells Promoted by Ginsenoside Rg1 In Vitro

“…2B). These observations demonstrated that HBASCs resemble other ASCs in terms of their morphology and proliferation capacity [26,28]. The HBASCs displayed positive staining for the mesenchymal surface markers CD29, CD44, CD49d, CD73, CD90, CD105 and HLA-ABC/FITC, as determined through flow cytometry analysis ( Table 3).…”

“…CD105, CD90 and CD44 are the three main positive markers used for MSCs, while CD34 is the most frequently reported negative marker. CD49d and CD31 have also been described as a positive marker and a negative marker, respectively [28].…”

Production of Human Dental Pulp Cells with a Medicinal Manufacturing Approach

“…As was previously shown for the isolation of human adipose or Wharton jelly stem/progenitor cells, minimal manipulation leads to higher safety and efficacy of cell production (15,16). In this study, we selected exclusively impacted third molars between Nolla developmental stages 5 (crown almost completed) and 7 (one third root completed) to minimize the risk of pulp tissue contamination with oral microorganisms and to avoid the cell stress that results from crown root mechanical separation during the pulp recovery from older teeth (4,14).…”

Isolation and Characterization of Human Mesenchymal Stromal Cell Subpopulations: Comparison of Bone Marrow and Adipose Tissue

“…We have chosen to deal with BM and AT as they represent the two main sources for MSC isolation in clinical trials [25]. AT seems to be an alternative source of MSC for cell-based therapy as some disadvantages are associated with BM [26,27] In our study, we demonstrated that MSC enrichment from a fresh sample was possible thanks to an appropriate cell selection. Importantly, the choice of the cell surface marker with which to proceed with the selection depended on the source of the sample as well as the therapeutical use of the MSC.…”