Stem cell engineering

The overall goal of the laboratory is to combine engineering principles with basic biological science to rationally understand the mechanisms governing cell behaviour. The research of the lab addresses fundamental and practical problems in the areas of stem cell engineering, cell therapy and development of in vitro models for functional healthy and diseased tissues.
We design and develop innovative technologies for the spatial-temporal control of stem cell culture at the micro-environmental scale. We are currently working on: methods of micro-fabrication; micro- and macro-bioreactor design; molecular cell biology; computational biology; microscopic mass transport modelling.

Human in vitro model

Skeletal and cardiac muscle play essential roles for human life but these complex tissues may be affected by a variety of pathologies which are in many cases invalidating or, in the worst case scenario, lethal. The elucidation of the intricate three dimensional structure-function relationship of skeletal and cardiac muscle tissues is certainly a critical issue for a deeper understanding of the physiological and the physiopathological processes in which they are involved.

In this context, we are focused on the development of in vitro models of human skeletal and cardiac muscle simulating the particular spatial arrangement of cells and extracellular matrix proteins, peculiar of their structure and function in vivo. Spatial control of the cells and substrate chemical and mechanical properties are obtained by micro-fabrication technologies and substrate engineering.

Human in vitro models could potentially represent a complementary tool bridging the gap between conventional cell culture, animal models and patients in the process of drugs or therapies development. Our research holds the potential to develop an in vitro model based on human artificial tissue for screening therapy and for the investigation of relevant molecular and biological mechanisms in a highthroughput fashion.

Figure 1. Human myoblast micro-patterning

Figure 1. Human myoblast micro-patterning
[click image to enlarge]

In vitro cardiac model

Cardiovascular diseases are the major cause of death in the modern society. At the moment, the available therapies are based on pharmacological treatments and organ transplantation, but they are not definitive. Regenerative medicine, based on the use of stem cells, is emerging as a promising approach for the treatment of failing hearts, but there is a great need for new laboratory models for studies on human cells and for conducting low cost preliminary testing prior to clinical trials. The choice of a good cell source is the key step for both regenerative medicine and laboratory model development. The possibility of reprogramming human adult cells (specialized to one precise cell type) to the embryonic state (capable of giving rise to all the cell type of the body and called induced pluripotent stem cells, iPS) has been recently discovered.
In this scenario, our research aims at developing a robust and efficient technology able to control and enhance the effectiveness of the reprogramming (from adult to embryonic state) and the programming process towards cells of the heart tissue, in order to provide cardiac cells to be used in clinical practice and regenerative medicine or for ad hoc models for drug screening and development.
This technology will be based on particular micro-bioreactors, called microfluidic platforms, able to ensure the fundamental requirements for the above mentioned processes: generation of extremely controlled and reproducible environments surrounding the cells for robust process development; accurate delivery of factors for high-efficiency reprogramming and differentiation to cardiac cells; micro-scale and multi-parametric assays for fast screening of experimental conditions; small volume devices, simple and versatile for low-cost experimentation.
The ultimate goal is thus to develop an integrated microfluidic platform able to assist the generation of patient specific human iPS cell line and their cardiac differentiation.

We also aim at studying the effects of the major pathological stimuli of an infarcted myocardium on the functional properties of embryonic-derived human cardiomyocytes. In this sight, we aim at realizing an in vitro model of infracted hearth, based on microstructured culture of human cardiomyocytes coupled with a microfluidic platform which allows the multi-parametric spatial-temporal simulation of the in vitro pathological environment.

Figure 2. Macro and micro-bioreactors for stem cell culture

Figure 2. Macro and micro-bioreactors for stem cell culture
[click image to enlarge]

In vitro skeletal muscle model

Duchenne Muscular Dystrophy is the most common, lethal, inherited disease of skeletal and cardiac muscles. Although several years have passed since the identification of the molecular defect involved in the Duchenne Dystrophy, effective therapies have not been developed yet. Our research efforts are aimed at producing in vitro human skeletal muscle myotubes exhibiting functional properties by a proper design of the in vitro artificial niche.
We aim at integrating tissue engineering tools and biological skills in order to reproduce the dominant physiological stimuli that guide myofiber formation in vivo; in particular, we culture dystrophic myoblasts on a hydrogel with mechanical properties resembling those of muscular tissue in vivo. Spatial alignment of the myoblasts is obtained by the micro-contact printing technique and electrical stimulation is coupled to the culture in order to mimic the physiological electrical signals of muscle tissue and study their functional activity.

Cell therapy

Cell therapy represents an appealing solution for the treatment of diseased muscle (both cardiac and skeletal muscle) or the reconstruction of muscle after severe injury. The success of innovative therapeutic strategies depend on the development of suitable technologies to engineer myogenic cells and biocompatible scaffolds.
In this perspective, we developed an injectable, biocompatible and biodegradable photo-polymerizable hydrogel able to maintain the myogenic potential of the cells and deliverable through minimally invasive techniques. To further improve the efficiency of cell delivery, survival, and integration with the host tissue we aim at developing injectable biomaterials functionalized with extracellular matrix proteins and with tuneable growth factors release rates.

Synoptic CV

Selected VIMM Publications

• Luni C, Feldman HC, Pozzobon M, De Coppi P, Meinhart CD, Elvassore N (2010) Microliter-bioreactor array with buoyancy-driven stirring for human hematopoietic stem cell culture. Biomicrofluidics 4:.
• Serena E, Zatti S, Reghelin E, Pasut A, Cimetta E, Elvassore N (2010) Soft substrates drive optimal differentiation of human healthy and dystrophic myotubes. Integr Biol (Camb) 2:193-201.
• Serena E, Figallo E, Tandon N, Cannizzaro C, Gerecht S, Elvassore N, Vunjak-Novakovic G (2009) Electrical stimulation of human embryonic stem cells: cardiac differentiation and the generation of reactive oxygen species. Exp. Cell Res. 315:3611-9.
• Flaibani M, Boldrin L, Cimetta E, Piccoli M, De Coppi P, Elvassore N (2009) Muscle differentiation and myotubes alignment is influenced by micropatterned surfaces and exogenous electrical stimulation. Tissue Eng Part A. 15:2447-57.
• Cimetta E, Figallo E, Cannizzaro C, Elvassore N, Vunjak-Novakovic G (2009) Micro-bioreactor arrays for controlling cellular environments: design principles for human embryonic stem cell applications. Methods 47:81-9.

Additional Publications

• Cimetta E, Pizzato S, Bollini S, Serena E, De Coppi P, Elvassore N (2009) Production of arrays of cardiac and skeletal muscle myofibers by micropatterning techniques on a soft substrate. 11:389-400.
• Serena E, Flaibani M, Carnio S, Boldrin L, Vitiello L, De Coppi P, Elvassore N (2008) Electrophysiologic stimulation improves myogenic potential of muscle precursor cells grown in a 3D collagen scaffold. Neurol. Res. 30:207-14.
• Cannizzaro C, Tandon N, Figallo E, Park H, Gerecht S, Radisic M, Elvassore N, Vunjak-Novakovic G (2007) Practical aspects of cardiac tissue engineering with electrical stimulation. Methods Mol. Med. 140:291-307.
• Callegari A, Bollini S, Iop L, Chiavegato A, Torregrossa G, Pozzobon M, Gerosa G, De Coppi P, Elvassore N, Sartore S (2007) Neovascularization induced by porous collagen scaffold implanted on intact and cryoinjured rat hearts. Biomaterials 28:5449-61.
• Cimetta E, Flaibani M, Mella M, Serena E, Boldrin L, De Coppi P, Elvassore N (2007) Enhancement of viability of muscle precursor cells on 3D scaffold in a perfusion bioreactor. Int J Artif Organs 30:415-28.
• Figallo E, Cannizzaro C, Gerecht S, Burdick JA, Langer R, Elvassore N, Vunjak-Novakovic G (2007) Micro-bioreactor array for controlling cellular microenvironments. 7:710-9.
• Figallo E, Flaibani M, Zavan B, Abatangelo G, Elvassore N (2007) Micropatterned biopolymer 3D scaffold for static and dynamic culture of human fibroblasts. Biotechnol. Prog. 23:210-6.

Selected Seminars

Contact

Nicola Elvassore Venetian Institute of Molecular Medicine Via Orus 2 35129 Padua — Italy