Abstract
Adipose-derived stromal/stem cells (ASCs) are currently being considered for clinical use for a number of indications. In order to develop standardized clinical protocols, it is paramount to have a full characterization of the stem cell preparations. The surface marker expression of ASCs has previously been characterized in multiple studies. However, most of these studies have provided a cross-sectional description of ASCs in either earlier or later passages. In this study, we evaluate the dynamic changes of 15 different surface molecules during culture. Using multichromatic flow cytometry, ASCs from three different donors each in passages 1, 2, 4, 6, and 8 were analyzed for their co-expression of markers associated with mesenchymal stem cells, wound healing, immune regulation, ASC markers, and differentiation capacity, respectively. We confirmed that at an early stage, ASC displayed a high heterogeneity with a plethora of subpopulations, which by culturing became more homogeneous. After a few passages, virtually all ASCs expressed CD29, CD166 and CD201, in addition to canonical markers CD73, CD90, and CD105. However, even at passage 8, there were several predominant lineages that differed with respect to the expression of CD34, CD200 and CD271. Although the significance of remaining subpopulations still needs to be elucidated, our results underscore the necessity to fully characterize ASCs prior to clinical use.
Highlights
Adipose-derived stromal/stem cells (ASCs) are a favorable type of mesenchymal stem cell (MSC)due to easy accessibility, high cell yield, and robust proliferative capacity [1,2]
We found that at an early stage, ASCs displayed a high found that at an early stage, ASCs displayed a high heterogeneity with a plethora of subpopulations, heterogeneity with became a plethora of subpopulations, which byofculturing becameexchange, homogenized due to which by culturing homogenized due to three ways re-distribution: converge, or three ways of re-distribution: exchange, converge, or diverge
Previous research has thatthat cultures are a mixture diverseof phenotypes, presumably research hasdemonstrated demonstrated cultures are a of mixture diverse phenotypes, with differentwith functionalities, and with a transcriptional which evolves along withevolves the expansion presumably different functionalities, and with apattern transcriptional pattern which along of thethe population donor variation and fat tissuevariation origin are additional contributing with expansion of theMoreover, population
Summary
Adipose-derived stromal/stem cells (ASCs) are a favorable type of mesenchymal stem cell (MSC)due to easy accessibility, high cell yield, and robust proliferative capacity [1,2]. ASCs have been recognized to be highly biologically active, with regenerative roles in tissue formation, homeostasis, and immune regulation [3,4,5]. These unique properties render ASCs attractive in scenarios where the traditional approaches fall short of the desired therapeutic outcome, such as for chronic wounds, diabetic ulcers, osteoarthritis, ischemic heart disease, or type 1 diabetes, just to mention a few important areas [6,7,8,9,10]. Provided that the discrete subpopulations can be linked with particular functionalities, it would be possible in the prospective cell-based protocols to tailor the desired properties to the clinical application, which would undoubtedly benefit the clinical outcome
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