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Evolution of ASC Immunophenotypical Subsets During Expansion In Vitro
Qiuyue Peng1, Hiva Alipour1, Simone Porsborg1
1Department of Health Science and Technology, Regenerative Medicine Group, Aalborg University, Fredrik Bajers Vej 3B, 9220 Aalborg, Denmark.
International Journal of Molecular Sciences
|February 26, 2020
Summary
Adipose-derived stromal/stem cells (ASCs) show dynamic surface marker changes during culture. Full characterization is essential before clinical application to understand cell heterogeneity and ensure safety and efficacy.
Area of Science:
- Cell Biology
- Regenerative Medicine
- Biotechnology
Background:
- Adipose-derived stromal/stem cells (ASCs) show promise for various clinical applications.
- Standardized clinical protocols require thorough characterization of ASC preparations.
- Previous studies often provided limited, cross-sectional data on ASC surface marker expression across passages.
Purpose of the Study:
- To investigate the dynamic changes in surface molecule expression of ASCs during extended in vitro culture.
- To analyze the heterogeneity and homogeneity of ASC populations across multiple passages (1-8).
- To identify key surface markers associated with ASCs, their differentiation, and immune functions over time.
Main Methods:
- Multichromatic flow cytometry was employed to analyze ASCs from three donors.
- ASCs were evaluated at passages 1, 2, 4, 6, and 8.
- Expression of 15 different surface molecules was assessed, including canonical MSC markers, immune regulatory markers, and differentiation markers.
Main Results:
- ASCs exhibited significant heterogeneity at early passages, which decreased with culturing, leading to a more homogeneous population.
- Canonical markers (CD73, CD90, CD105) and others (CD29, CD166, CD201) became widely expressed by passage 8.
- Distinct subpopulations persisted at passage 8, differing in the expression of CD34, CD200, and CD271.
Conclusions:
- ASC surface marker expression is dynamic and changes significantly with in vitro culture duration.
- While ASCs become more homogeneous, specific subpopulations with unique marker profiles remain even at later passages.
- Comprehensive characterization of ASCs across passages is crucial for their safe and effective clinical translation.
