Adipose derived stem cells (ASCs) hold great promise for clinical applications such as soft tissue regeneration and for in vitro tissue models and are notably easy to derive in large numbers. Specifically, ASCs provide an advantage for in vitro models of adipose tissue, where they can be employed as tissue specific cells and for patient specific models. However, ASC in vitro expansion may unintentionally reduce adipogenic capacity due to the stiffness of tissue culture plastic (TCPS).
Here, we expanded freshly isolated ASCs on soft and stiff substrates for 4 passages before adipogenic differentiation. At the last passage we swapped the substrate from stiff to soft, or soft to stiff to determine if short term exposure to a different substrate altered adipogenic capacity.
Expansion on stiff substrates reduced adipogenic capacity by 50% which was not rescued by swapping to a soft substrate for the last passage. Stiff substrates had greater nuclear area and gene expression of nesprin-2, a protein that mediates the tension of the nuclear envelope by tethering it to the actin cytoskeleton. Upon swapping to a soft substrate, the nuclear area was reduced but nesprin-2 levels did not fully recover, which differentially regulated cell commitment transcriptional factors.
Therefore, in vitro expansion on stiff substrates must be carefully considered when the end-goal of the expansion is for adipose tissue or soft tissue applications.
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The authors would like to acknowledge funding support from Dr. Bellas’s startup funds from Temple University College of Engineering and the NIH NIDDK Diabetic Complications Consortium DK07616 and DK115255 grants (to E.B.) for their financial support toward this project. The authors would also like to thank the Temple University School of Medicine’s Cardiovascular Institute for use of their QPCR instrument and Drs. Andrew Gassman, M.D. and George Taylor, M.D. for procuring the post-surgical specimens.
Conflict of interest
Author Berger, Author Anvari and Author Bellas declare that they have no conflict of interest.
All human subjects research was carried out in accordance with institutional guidelines and approved by the Temple University Intuitional Review Board. No animals studies were performed.
Associate Editor Michael R. King oversaw the review of this article.
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This article is part of the 2021 CMBE Young Innovators special issue.
Evangelia Bellas is an Assistant Professor in the Department of Bioengineering at Temple University. Prior to joining Temple University, Dr. Bellas was a postdoctoral fellow in Biomedical Engineering at Boston University and Bioengineering at University of Pennsylvania under the mentorship of Dr. Christopher Chen where she developed 3D in vitro adipose tissue disease models. She received her Ph.D. in Biomedical Engineering at Tufts University mentored by Dr. David Kaplan. Her Ph.D. research focused on developing long-term volume stable silk biomaterials for soft tissue regeneration. This work resulted in 2 patents and a start-up. Before starting her Ph.D., Dr. Bellas was at Massachusetts Institute of Technology under the supervision of Drs. Robert Langer and Daniel Kohane, where she worked on biomaterial, drug delivery solutions for prevention of peritoneal adhesions and controlled release formulations for long-term pain management. Her current research focuses on the development of fat-on-chip and (dys)functional adipose tissue models to study how vascularization and interactions with the microenvironment impact tissue health and function and funded by NIH, NASA, NSF. Dr. Bellas is active in diversity, equity and inclusion efforts and currently serves as the Biomedical Engineering Society’s Diversity Committee Chair.
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Berger, A.J., Anvari, G. & Bellas, E. Mechanical Memory Impairs Adipose-Derived Stem Cell (ASC) Adipogenic Capacity After Long-Term In Vitro Expansion. Cel. Mol. Bioeng. 14, 397–408 (2021). https://doi.org/10.1007/s12195-021-00705-9
- Stem cell fate
- Biophysical cues
- Substrate rigidity