Proliferative and phenotypical characteristics of human adipose tissue-derived stem cells: comparison of Ficoll gradient centrifugation and red blood cell lysis buffer treatment isolation methods

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Abstract

Background: Adult human subcutaneous adipose tissue harbours a multipotent stem cell population, the so- called human adipose tissue-derived stromal cells (AT-MSCs). These cells are able to differentiate in vitro into various cell types and possess immunomodulatory features. Yet, procedures to obtain AT-MSCs can vary significantly. The two most extensively used AT-MSC isolation techniques are (i) density gradient centrifugation using Ficoll and (ii) Red Blood Cell (RBC) lysis buffer treatment of the stromal vascular fraction (SVF). In the context of potential clinical cell therapy, the stem cell yield after isolation and upon consecutive passages as well as the purity of the obtained cell population are of utmost importance.
Methods: We investigated the expansion capacity and purity of AT-MSCs isolated by both isolation procedures immediately after isolation and upon consecutive passages. We also investigated possible isolation-dependent differences in their expression of immune inhibitory factors and cell adhesion molecules (CAM).
Results: We found that RBC lysis buffer treatment is a more robust and easier method to isolate AT-MSCs than density gradient fractionation. Yet, the resulting AT-MSC-RBC population contains a significantly higher amount of CD34+ cells, particularly during the first passages after plating. From passage 4 onwards, no significant differences could be observed between both populations with respect to the immunophenotype, expansion capacity and expression of immune inhibitory factors and CAMs.
Discussion: Our data shows that RBC lysis buffer treatment may be a good alternative to density fractionation as being a faster, more robust and easier method to isolate AT-MSCs with biologically preserved characteristics.
Original languageEnglish
Pages (from-to)1220-1228
Number of pages9
JournalCytotherapy
Volume16
Issue number9
Publication statusPublished - 2014

Keywords

  • adipose tissue
  • mesenchymal stromal cell
  • stromal vascular fraction
  • Ficoll
  • red blood cell lysis
  • density gradient centrifugation
  • adult stem cell

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