Mechanically tuned 3 dimensional hydrogels support human mammary fibroblast growth and viability

BMC Cell Biology

Research Article

Jennifer Peyling Wang¹  Hana Park²  Abigail Hielscher²  Catlyn Thigpen²  Kathryn Woods³ 

[1] Anschutz Medical Campus Skaggs School of Pharmacy, University of Colorado, 12850 E. Montview Blvd, 80011, Aurora, CO, USA;Department of Biomedical Sciences, Georgia Philadelphia College of Osteopathic Medicine, 30024, Suwanee, GA, USA;Department of Pathology and Microbiology, University of Nebraska Medical Center, 985900 Nebraska Medical Center, 68198, Omaha, NE, USA;

关键词: 3 dimensional hydrogels;    Microbial transglutaminase;    Myofibroblasts;    Human mammary fibroblasts;    Extracellular matrix;   

DOI  :  10.1186/s12860-017-0151-y

 received in 2017-04-13, accepted in 2017-12-07,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundCarcinoma associated fibroblasts (CAFs or myofibroblasts) are activated fibroblasts which participate in breast tumor growth, angiogenesis, invasion, metastasis and therapy resistance. As such, recent efforts have been directed toward understanding the factors responsible for activation of the phenotype. In this study, we have investigated how changes in the mechanical stiffness of a 3D hydrogel alter the behavior and myofibroblast-like properties of human mammary fibroblasts (HMFs).ResultsHere, we utilized microbial transglutaminase (mTG) to mechanically tune the stiffness of gelatin hydrogels and used rheology to show that increasing concentrations mTG resulted in hydrogels with greater elastic moduli (G’). Upon encapsulation of HMFs in 200 (compliant), 300 (moderate) and 1100 Pa (stiff) mTG hydrogels, it was found that the HMFs remained viable and proliferated over the 7 day culture period. Specifically, rates of proliferation were greatest for HMFs in moderate hydrogels. Regarding morphology, HMFs in compliant and moderate hydrogels exhibited a spindle-like morphology while HMFs in stiff hydrogels exhibited a rounded morphology with several large cellular protrusions. Quantification of cell morphology revealed that HMFs cultured in all mTG hydrogels overall assumed a more elongated phenotype over time in culture; however, few significant differences in morphology were observed between HMFs in each of the hydrogel conditions. To determine whether matrix stiffness upregulated expression of ECM and myofibroblast markers, western blot was performed on HMFs in compliant, moderate and stiff hydrogels. It was found that ECM and myofibroblast proteins varied in expression during both the culture period and according to matrix stiffness with no clear correlation between matrix stiffness and a myofibroblast phenotype. Finally, TGF-β levels were quantified in the conditioned media from HMFs in compliant, moderate and stiff hydrogels. TGF-β was significantly greater for HMFs encapsulated in stiff hydrogels.ConclusionsOverall, these results show that while HMFs are viable and proliferate in mTG hydrogels, increasing matrix stiffness of mTG gelatin hydrogels doesn’t support a robust myofibroblast phenotype from HMFs. These results have important implications for further understanding how modulating 3D matrix stiffness affects fibroblast morphology and activation into a myofibroblast phenotype.

【 授权许可】

CC BY   
© The Author(s). 2017