Baaijens, FPT Frank

Boonen, KJM Kristel

Post, MJ Mark

Repository TU/e

Mimicking the satellite cell niche

Boonen, K.J.M.

Baaijens, F.P.T.

Post, M.J.

Pure OAI Repository

 Mimicking the satellite cell nicheCitation for published version (APA):Boonen, K. J. M., Baaijens, F. P. T., & Post, M. J. (2007). Mimicking the satellite cell niche. Poster sessionpresented at Mate Poster Award 2007 : 12th Annual Poster Contest.Document status and date:Published: 01/01/2007Document Version:Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers)Please check the document version of this publication:• A submitted manuscript is the version of the article upon submission and before peer-review. There can beimportant differences between the submitted version and the official published version of record. Peopleinterested in the research are advised to contact the author for the final version of the publication, or visit theDOI to the publisher's website.• The final author version and the galley proof are versions of the publication after peer review.• The final published version features the final layout of the paper including the volume, issue and pagenumbers.Link to publicationGeneral rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright ownersand it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.            • Users may download and print one copy of any publication from the public portal for the purpose of private study or research.            • You may not further distribute the material or use it for any profit-making activity or commercial gain            • You may freely distribute the URL identifying the publication in the public portal.If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, pleasefollow below link for the End User Agreement:www.tue.nl/taverneTake down policyIf you believe that this document breaches copyright please contact us at:openaccess@tue.nlproviding details and we will investigate your claim.Download date: 04. Oct. 2023IntroductionBecause of their effectiveness in regenerating large muscle defects in vivo, we investigate the use of satellite cells derived from muscle biopsies as a cell source for muscle tissue engineering. Despite their in vivo potency, however, their proliferative and myogenic capacity in vitro has been disappointing.1We therefore concentrate on optimizing culture conditions by mimicking the natural niche environment of these cells. The elasticity of the environment is an important aspect of this niche and has been shown to influence differentiation pathways of stem cells2. We hope to improve the in vitro capacities of satellite cells by varying substrate elasticity. The potential for myogenesis is evaluated from morphological and functional parameters. The model system used in these experiments provides for high-throughput and well-controlled measurements and can ultimately be expanded to 3D tissue culture conditions using biomaterials and/or different cell types. Material and methodsTibialis Anterior (TA) muscles from Swiss White mice are digested in collagenase and triturated to release single fibres from which satellite cells are liberated with a 19G needle. The satellite cells are plated on  MatrigelTM-coated polyacrylamide(PA) gels (Figure 1) with elastic moduli (E) ranging from 1.4 to72 kPa on top of 13 mm ø coverslips and cultured in plating medium (DMEM advanced, 10% HS, 20% FBS, 0.5% CEE, 1% pen/strep, 1% L-glut.). Results• Satellite cells proliferate on all substrates • Fusion first takes place on 17 kPa, which mimics physiological elasticity of striated muscle. • Subsequently, satellite cells fuse on 8.1 and 35 kPa• Eventually fusion takes place on all E-moduliConclusionsTiming of differentiation of satellite cells seems to be dependent on the elasticity of their environment. Fusion first takes place on physiological elasticity of striated muscle, subsequently at the elastic moduli closest to this physiological value and lastly at the outer values.Overall, we have developed a robust tissue culture system, that gives us the possibility to pursue long term proliferation of satellite cell derived progenitor cells and evaluate their proliferative and differentiative capacities. In the future, different biochemical and biophysical cues will be used to influence and optimize these processes. References:[1] Collins, C. A., I. Olsen, et al. (2005). "Stem Cell Function, Self-Renewal, and Behavioral Heterogeneity of Cells from the Adult Muscle Satellite Cell Niche." Cell 122(2): 289-301.[2] Engler, A. J., S. Sen, et al. (2006). "Matrix elasticity directs stem cell lineage specification." Cell 126(4): 677-89.Mimicking the satellite cell nicheK.J.M. Boonen, F.P.T. Baaijens, M.J. PostEindhoven University of Technology, Department of Biomedical EngineeringFigure 1: Creation of coated PA gels on top of coverslipsDay 16DAPI/Actin/desmin1.4 kPaDay 15Fusion all E-moduliDay 12Fusion 8.1 & 35 kPaDay 9Fusion 17 kPaDay 7Big colonies72 kPa35 kPa17 kPa8.1 kPaFigure 2: Proliferation and differentiation of satellite cells on substrates with                   varying elasticity

English

 Mimicking the satellite cell nicheCitation for published version (APA):Boonen, K. J. M., Baaijens, F. P. T., & Post, M. J. (2007). Mimicking the satellite cell niche. Poster sessionpresented at Mate Poster Award 2007 : 12th Annual Poster Contest.Document status and date:Published: 01/01/2007Document Version:Publisher’s PDF, also known as Version of Record (includes final page, issue and volume numbers)Please check the document version of this publication:• A submitted manuscript is the version of the article upon submission and before peer-review. There can beimportant differences between the submitted version and the official published version of record. Peopleinterested in the research are advised to contact the author for the final version of the publication, or visit theDOI to the publisher's website.• The final author version and the galley proof are versions of the publication after peer review.• The final published version features the final layout of the paper including the volume, issue and pagenumbers.Link to publicationGeneral rightsCopyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright ownersand it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights.            • Users may download and print one copy of any publication from the public portal for the purpose of private study or research.            • You may not further distribute the material or use it for any profit-making activity or commercial gain            • You may freely distribute the URL identifying the publication in the public portal.If the publication is distributed under the terms of Article 25fa of the Dutch Copyright Act, indicated by the “Taverne” license above, pleasefollow below link for the End User Agreement:www.tue.nl/taverneTake down policyIf you believe that this document breaches copyright please contact us at:openaccess@tue.nlproviding details and we will investigate your claim.Download date: 17. Nov. 2023IntroductionBecause of their effectiveness in regenerating large muscle defects in vivo, we investigate the use of satellite cells derived from muscle biopsies as a cell source for muscle tissue engineering. Despite their in vivo potency, however, their proliferative and myogenic capacity in vitro has been disappointing.1We therefore concentrate on optimizing culture conditions by mimicking the natural niche environment of these cells. The elasticity of the environment is an important aspect of this niche and has been shown to influence differentiation pathways of stem cells2. We hope to improve the in vitro capacities of satellite cells by varying substrate elasticity. The potential for myogenesis is evaluated from morphological and functional parameters. The model system used in these experiments provides for high-throughput and well-controlled measurements and can ultimately be expanded to 3D tissue culture conditions using biomaterials and/or different cell types. Material and methodsTibialis Anterior (TA) muscles from Swiss White mice are digested in collagenase and triturated to release single fibres from which satellite cells are liberated with a 19G needle. The satellite cells are plated on  MatrigelTM-coated polyacrylamide(PA) gels (Figure 1) with elastic moduli (E) ranging from 1.4 to72 kPa on top of 13 mm ø coverslips and cultured in plating medium (DMEM advanced, 10% HS, 20% FBS, 0.5% CEE, 1% pen/strep, 1% L-glut.). Results• Satellite cells proliferate on all substrates • Fusion first takes place on 17 kPa, which mimics physiological elasticity of striated muscle. • Subsequently, satellite cells fuse on 8.1 and 35 kPa• Eventually fusion takes place on all E-moduliConclusionsTiming of differentiation of satellite cells seems to be dependent on the elasticity of their environment. Fusion first takes place on physiological elasticity of striated muscle, subsequently at the elastic moduli closest to this physiological value and lastly at the outer values.Overall, we have developed a robust tissue culture system, that gives us the possibility to pursue long term proliferation of satellite cell derived progenitor cells and evaluate their proliferative and differentiative capacities. In the future, different biochemical and biophysical cues will be used to influence and optimize these processes. References:[1] Collins, C. A., I. Olsen, et al. (2005). "Stem Cell Function, Self-Renewal, and Behavioral Heterogeneity of Cells from the Adult Muscle Satellite Cell Niche." Cell 122(2): 289-301.[2] Engler, A. J., S. Sen, et al. (2006). "Matrix elasticity directs stem cell lineage specification." Cell 126(4): 677-89.Mimicking the satellite cell nicheK.J.M. Boonen, F.P.T. Baaijens, M.J. PostEindhoven University of Technology, Department of Biomedical EngineeringFigure 1: Creation of coated PA gels on top of coverslipsDay 16DAPI/Actin/desmin1.4 kPaDay 15Fusion all E-moduliDay 12Fusion 8.1 & 35 kPaDay 9Fusion 17 kPaDay 7Big colonies72 kPa35 kPa17 kPa8.1 kPaFigure 2: Proliferation and differentiation of satellite cells on substrates with                   varying elasticity

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Mimicking the satellite cell niche

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