Human Pluripotent Stem Cells (hPSCs) hold great promise for cell therapy and tissue engineering, as well as drug screening. For a clinically valid development of stem cell-based therapies some biological and engineering challenges still need to be overcome, such as the design of engineered cell culture microenvironment that support hPSCs proliferation while maintaining their pluripotency. In this study a novel composite synthetic scaffold was designed, inspired by the overall structure of tissue extracellular matrix (ECM), and the short-time expansion and self-renewal of human Embryonic Stem Cells (hESCs) and human Induced Pluripotent Stem Cells (hiPSCs) was investigated in view of potential application of the materials as scaffolds for stem cel-mediated tissue engineering applications. The scaffold was composed by an arg-gly-asp (RGD)-mimic polyamidoamine hydrogel with embedded poly-L-lactic acid (PLLA) mat of continuous electrospun nanofibers with average diameter 570 ± 170 nm, mimicking the gel and fibrous components of ECM, respectively. The biomimetic properties and the softness of the hydrogel component were therefore combined with the strength of the nanofibrous PLLA mat. Strong matrix-fiber adhesion was achieved by N2 atmospheric pressure non-equilibrium plasma treatment of the PLLA mat. The scaffolds were characterized for their swelling and degradation behavior and their mechanical properties were investigated. Biological studies demonstrated that the scaffolds supported short term self-renewal of Human Pluripotent Stem cells in feeder-free conditions and that the cells fully retained stemness for at least 7 days. The novel composites developed in this work are entirely synthetic and they can be obtained by standardized procedures. Moreover, their mechanical properties, swelling and degradation behavior in aqueous media can be easily tuned by tailoring hydrogel crosslinking degree. Prospectively, their chemical structure is suitable to covalently incorporate bioactive molecules in order to specifically modulate the signaling pathways of hiPSCs.

Poly- L-lactic acid nanofiber–polyamidoamine hydrogel composites: preparation, properties, and preliminary evaluation as scaffolds for human pluripotent stem cell culturing / M. Letizia Focarete, C. Gualandi, N. Bloise, N. Mauro, P. Ferruti, A.G. Manfredi, M. Sampaolesi, A. Liguori, R. Laurita, M. Gherardi, V. Colombo, L. Visai, E. Ranucci. ((Intervento presentato al convegno Milan Polymer Days tenutosi a Milano nel 2017.

Poly- L-lactic acid nanofiber–polyamidoamine hydrogel composites: preparation, properties, and preliminary evaluation as scaffolds for human pluripotent stem cell culturing

P. Ferruti;A.G. Manfredi;E. Ranucci
2017

Abstract

Human Pluripotent Stem Cells (hPSCs) hold great promise for cell therapy and tissue engineering, as well as drug screening. For a clinically valid development of stem cell-based therapies some biological and engineering challenges still need to be overcome, such as the design of engineered cell culture microenvironment that support hPSCs proliferation while maintaining their pluripotency. In this study a novel composite synthetic scaffold was designed, inspired by the overall structure of tissue extracellular matrix (ECM), and the short-time expansion and self-renewal of human Embryonic Stem Cells (hESCs) and human Induced Pluripotent Stem Cells (hiPSCs) was investigated in view of potential application of the materials as scaffolds for stem cel-mediated tissue engineering applications. The scaffold was composed by an arg-gly-asp (RGD)-mimic polyamidoamine hydrogel with embedded poly-L-lactic acid (PLLA) mat of continuous electrospun nanofibers with average diameter 570 ± 170 nm, mimicking the gel and fibrous components of ECM, respectively. The biomimetic properties and the softness of the hydrogel component were therefore combined with the strength of the nanofibrous PLLA mat. Strong matrix-fiber adhesion was achieved by N2 atmospheric pressure non-equilibrium plasma treatment of the PLLA mat. The scaffolds were characterized for their swelling and degradation behavior and their mechanical properties were investigated. Biological studies demonstrated that the scaffolds supported short term self-renewal of Human Pluripotent Stem cells in feeder-free conditions and that the cells fully retained stemness for at least 7 days. The novel composites developed in this work are entirely synthetic and they can be obtained by standardized procedures. Moreover, their mechanical properties, swelling and degradation behavior in aqueous media can be easily tuned by tailoring hydrogel crosslinking degree. Prospectively, their chemical structure is suitable to covalently incorporate bioactive molecules in order to specifically modulate the signaling pathways of hiPSCs.
15-feb-2017
Tissue engineering; bioactive and biocompatible polymers
Settore CHIM/04 - Chimica Industriale
Università di Milano; CNR-ISMAC; INSTM
http://www.mipol.unimi.it/2017/index.html
Poly- L-lactic acid nanofiber–polyamidoamine hydrogel composites: preparation, properties, and preliminary evaluation as scaffolds for human pluripotent stem cell culturing / M. Letizia Focarete, C. Gualandi, N. Bloise, N. Mauro, P. Ferruti, A.G. Manfredi, M. Sampaolesi, A. Liguori, R. Laurita, M. Gherardi, V. Colombo, L. Visai, E. Ranucci. ((Intervento presentato al convegno Milan Polymer Days tenutosi a Milano nel 2017.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/515468
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