SAVVY

Self-assembled virus-like vectors for stem cell phenotyping

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 Obiettivo del progetto (Objective)

'The Self-Assembled Virus-like Vectors for Stem Cell Phenotyping (SAVVY) project relies on hierarchical, multi-scale assembly of intrinsically dissimilar nanoparticles to develop novel types of multifunctional Raman probes for analysis and phenotyping of heterogeneous stem cell populations. Our project will address a large unmet need, as stem cells have great potential for a broad range of therapeutic and biotechnological applications. Characterization and sorting of heterogeneous stem cell populations has been intrinsically hampered by (1) lack of specific antibodies, (2) need for fluorescence markers, (3) low concentration of stem cells, (4) low efficiencies/high costs. Our technology will use a fundamentally different approach that (1) does not require antibodies, aptamers, or biomarkers, (2) is fluorescence-label free, and (3) is scalable at acceptable cost. The approach uses intrinsic differences in the composition of membranes of cells to distinguish cell populations. These differences will be detect by SERS and analysed through multicomponent analysis. We have combined the necessary expertise to tackle this challenge: Stellacci has developed rippled nanoparticles that specifically interact with and adhere to cell membranes (analogues to cell penetrating peptides). Lahann has developed bicompartmental Janus polymer particles that have already been surface-modified with rippled particles and integrate specifically in the cell membrane (analogues to viruses). Liz-Marzan has developed highly Raman-active nanoparticles and has demonstrated their selectivity and specificity in SERS experiments. These Raman probes will be loaded into the synthetic viruses to enable membrane fingerprinting. Stevens has developed a Bioinformatics platform for fingerprinting of stem cell populations using cluster analysis algorithms. The effort will be joined by two SMEs, ChipShop and OMT, that will be able to develop the necessary microfluidic and Raman detection hardware.'

Introduzione (Teaser)

Stem cells have a great potential for many therapeutic and biotechnological applications. An EU-funded project is developing a new approach to sort stem cell populations.

Descrizione progetto (Article)

Characterisation and sorting of stem cell populations is typically performed using flow cytometry. Several factors limit existing sorting technologies, such as lack of specific antibodies, low concentration of stem cells and a need for fluorescence markers.

The http://savvyproject.eu/ (SAVVY) (Self-assembled virus-like vectors for stem cell phenotyping) project is developing a different particle-based approach to cell sorting. The main idea is to use the molecular signatures of heterogeneous stem cell populations to enable their effective separation.

Running for three years to end-2015, the project uses intrinsic differences in the composition of cell membranes to distinguish and ultimately sort stem cell populations. SAVVY's approach does not require antibodies, aptamers or other biomarkers, and it is label-free and scalable at an acceptable cost.

The project relies on multi-scale assembly of intrinsically dissimilar nanoparticles (NPs) to develop novel types of multifunctional Raman probes. The completed live cell sorting system will incorporate NP-based signal enhancers along with Raman microspectroscopy in an integrated microfluidic cell sorter.

To date, the project has produced bicompartmental polymer particles, modified with gold NPs. These SAVVY reporters are currently under investigation concerning their interactions with cell surfaces.

At the same time, a database of Raman signatures of cells is under development. Scientists are performing Raman mapping of human and mouse embryonic stem cells using visible and near-infrared lasers. A first iteration of a Raman signature library was built.

In addition, SAVVY is looking for the surface-enhanced Raman scattering (SERS) signatures of cells. To obtain a model system for the SERS-based phenotyping, scientists applied gold nanostars, functionalised with cell-adhesion peptides, to human neural stem cells and human neurons.

This new approach to phenotyping and sorting will be useful for stem cell-based therapies, tissue regeneration and novel medical technologies. Once validated for stem cell phenotyping, the SAVVY technology may lead to novel diagnostics tools applicable to a number of diseases.