Dynamic surfaces to mimic mesenchymal stem cell niche functions

Lead Research Organisation: University of Glasgow
Department Name: College of Medical, Veterinary &Life Sci

Abstract

We live in an ageing society and we are outliving the useful lives of our bodies. Structural components suffer with arthritis or osteoporosis and organs provide reduced efficiency and can become damaged or diseased through degenerative processes. We live at an exciting point in history where we all have the expectation that unlocking the potential of stem cells will help with these urgent regenerative demands. Embryonic stem cells remain locked in ethical debate, however, and also have clinical issues associated with their use (including lack of immune privilege, which can cause adverse immune reactions, and the possibility of teratoma formation, which is a type of cancer ). Adult stem cells provide an alternate route with mesenchymal stem cells from, for example, bone marrow (obtained by e.g. marrow donation) or fat tissue (obtained by e.g. liposuction) providing an attractive, autologous (i.e. from the patient) source of multipotent cells.
A major hurdle with adult stem cells is their rapid and spontaneous differentiation during standard culture in the lab (i.e. out of the body they rapidly stop acting as stem cells). Current cell culture materials were developed before our understanding of stem cells had matured and were designed to grow mature cell types (such as fibroblasts) or cell lines (such as HeLa cells). Thus, we are currently lacking good platforms for autologous stem cell growth.
In the last few years, researchers, including ourselves, have understood that MSC growth and differentiation is controlled by the way cells adhere to materials and consistent 'rules' are starting to emerge. Developments in materials science have put forwards surfaces that are either favourable for MSC growth or good for differentiation, however, but that cannot control both.
In our bodies, stem cells reside in specialised locations (called 'niches') that control their growth to allow a supply of stem cells to be present in tissues throughout our lives and also regulate differentiation in response to tissue demand. It is, again, considered that cell adhesion is key to the niche regulation of stem cells.
Here, we will develop highly novel materials that initially support the growth (multiplication) of multipotent MSCs, which can then be switched under user control to turn on the desired type of differentiation, to generate the mature 'functional' cells of the body. To do this, we will use enzymes (biological catalysts) to cleave the self-renewal surface (this will be made by use of adhesion controlling chemistry and use of nanoscale spatial information i.e. small chemical patterns) and reveal the underlying differentiation surface (different chemistries to control differential adhesion, and hence drive stem cell fate). Such enzymes can be simply added by the user to the cell media (their food). We will then go further and place the switch under cell control. As cells become dense in a culture (near confluence) their protein (and hence enzyme) profile changes and we will exploit this to find enzymes that can perform the switch from a growth-promoting substrate to a differentiation-inducing substrate, only after the cells have grown to large numbers.
This technology will act as a platform for MSC growth and differentiation. It will be dynamic, as their natural niche is dynamic, and it will be an important step in the development of production of autologous cells with therapeutic potential.

Technical Summary

The stem cell niche acts to control stem cell growth (control self-renewal) and differentiation in a dynamic manner. Current developments in materials science, focused on a key regenerative stem cell type, the mesenchymal stem cell, have allowed us to regulate both self-renewal (to allow us to grow multipotent populations by preventing phenotypical drift to e.g. fibroblasts) and to target differentiation to desired functional cell types. The materials do this by controlling cell adhesion and intracellular tension important for regulation of biochemistry and transcription. However, the materials are designed either for growth or for differentiation and lack the flexibility to support both stem cell functions.
We have developed enzyme switchable surfaces that can be used to control cell adhesion in an on-demand manner and will exploit this technology to develop dynamic materials for stem cell culture. In the proposal, we will create an initial surface that control adhesion to promote self-renewal and hence growth as multipotent stem cells allowing us to achieve large numbers of true stem cells. This will be achieved by controlling the length of polyethyleneglycol (PEG) chains that block cells from adhering to the underlying differentiation surface (e.g. RGD, t-butyl, single peptides). Enzymatic cleavage of the PEG chains will then expose the differentiation surface to the mesenchymal stem cells on demand. Hence we will have fabricated surfaces that allow the cells to grow to large numbers and then change to functional cells in a controllable manner mimicking niche dynamics.
Finally, we will explore the possibility of using secreated enzymes (e.g. matrix metalloproteases) which have changes expression profiles in dense cultures to switch the surfaces as the cells grow more confluent (i.e. the enzymatic switch would automatically happen when enough stem cells were present).

Planned Impact

Initially impact will be academic as this is a very new concept. However, it is recognized that a step change is required to facilitate therapeutic use of stem cells as traditional cell culture plastics are not suitable and soluble factor chemistry is proving complex and can add artefact and so this research is vital. The use of niche biomimicry (stem cells responding on demand) will capture the imagination of science in general and we will target high-impact journals to ensure maximum dissemination to the community.

It is noteworthy that the applicants have strong track record in taking fundamental research observations towards commercialisation through spin-out and licensing. We will, again, look to protect IP and then exploit this towards next generation stem cell culture products. Such products will impact on cell culture industry as new products for stem cell growth are made, biotechnology industry will benefit as capacity for stem cell growth increases and autologous stem cell therapies become available. Tissue engineering will benefit as lots of high-quality cells from small isolations can be achieved and incorporated in their scaffold materials. Finally, clinicians and the public will benefit from therapies enabled by our platform technologies. This exploitation will be ongoing through the grant.

Thus, non-academic beneficiaries will include:
1) Industry - the development of new materials will fuel stem cell research. Stem cells themselves can now be purchased from catalogues and we will provide new consumables for purchase alongside the cells to aid research.
2) Clinic - most clinicians now accept that new biomaterials coupled to stem cells and nanotechnology will be important in the medium-term future. We believe it is pivotal for clinicians to engage with early-stage research to inform a realistic compromise between material function and clinical application.
3) Patients - the public are interested in (and actually expect) the delivery of stem cell therapies and our surfaces hold the potential to help deliver these therapies.
4) Public - through our public outreach in schools (Dalby has close ties with Lenzie Academy). We will inspire students to consider a career in research and academia. There is a poor understanding amongst school children taking career decision about what academics do beyond teaching and the best students choose medicine as a default career. However, basic science can be just as rewarding, if not more so, and school leavers should be aware of the possibility to help change medicine through academic research. It is, we believe, encumbent on us to stimulate, inform and inspire the next generation as an informed public base will lead to an informed science culture. Dalby had participated in stem cell debates with the public (for BBSRC, EPSRC and Royal Society of Edinburgh) and will continue this work.

To support this impact we will use our surgical links with Mr Meek through the Glasgow Orthopaedic Research Initiative and also with Prof Hart for plastic and reconstructive surgical links. They are both keen to engage with fundamental science to help facilitate translation.

Publications

10 25 50
 
Description We created an approach to make surfaces that respond dynamically to enzymes recreated by mesenchymal stems cells (MSCs). Stem cell care dynamic and to fully understand these cells with large regenerative potential we need to think better about how we culture them to mimic the dynamic nature of their niche environment. The surfaces created in the application allowed cells to growth and then differentiate in a. more niche-like manner and represent a future of stem cell substrates.
Exploitation Route The outcomes have helped us move towards developing better stem cell culture environments and we are now using this knowledge to create supportive environments for other stem cells and to produced environments where MSC niche phenotypes, such as immunomodulation can be enhanced. This will support future therapies such as transplantation.
Sectors Healthcare

 
Description The findings have been used widely in outreach and public presentations. We have also taken the knowledge forwards into the design of new niches to support MSC immunomodulation and haematopoetic stem cell maintenance 9papers in prep). The work performed in this grant will be fundamental to the next generation of MSC support materials.
First Year Of Impact 2014
Sector Healthcare
Impact Types Societal

 
Description Glasgow Orthopaedic Research Initiative
Geographic Reach National 
Policy Influence Type Influenced training of practitioners or researchers
 
Description Healickick (EU grant) translational day
Geographic Reach Multiple continents/international 
Policy Influence Type Influenced training of practitioners or researchers
Impact Preparing surgeons for the technologies and getting advice how to best present the technologies in product form
 
Description 'EPSRC and SFI Centre for Doctoral Training in Engineered Tissues for Discovery, Industry and Medicine
Amount £7,013,578 (GBP)
Funding ID EP/S02347X/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 05/2019 
End 12/2027
 
Description Engineering growth factor microenvironments - a new therapeutic paradigm for regenerative medicine
Amount £3,661,144 (GBP)
Funding ID EP/P001114/1 
Organisation Engineering and Physical Sciences Research Council (EPSRC) 
Sector Public
Country United Kingdom
Start 09/2016 
End 09/2021
 
Title Switchable surfaces for stem cells 
Description We have developed a surface that can either be user switched or cell switched to change during culture to allow firstly mesenchymal stem cell growth and then stem cell differentiation to osteoblasts 
Type Of Material Model of mechanisms or symptoms - in vitro 
Provided To Others? No  
Impact Non yet, but the technology used as the switch is being used in a wide number of systems with a range of biological collaborators. 
 
Title Dynamic surfaces for the study of mesenchymal stem cell growth through adhesion regulation 
Description  
Type Of Material Database/Collection of data 
Year Produced 2016 
Provided To Others? Yes  
 
Title Improved hyaline cartilage phenotype from pericytes differentiated using tunable peptide hydrogels 
Description  
Type Of Material Database/Collection of data 
Year Produced 2016 
Provided To Others? Yes  
 
Title Using tunable hydrogels for selection of lineage guiding metabolites in stem cell cultures 
Description  
Type Of Material Database/Collection of data 
Year Produced 2015 
Provided To Others? Yes  
 
Description COllaboration with Atelerix 
Organisation Atelerix
Country United Kingdom 
Sector Private 
PI Contribution Thi sis a joint PhD project in which we are designing in vitro stem cell niches
Collaborator Contribution Provision and tailoring of hydrogel materials
Impact Early collaboration
Start Year 2021
 
Description Collaboration for soilid phase peptide synthesis 
Organisation City University of New York (CUNY)
Country United States 
Sector Academic/University 
PI Contribution We worked with this group to create the peptide surfaces used in the project - we did the cell biology analysis of the peptide surfaces
Collaborator Contribution They made the surfaces and characterised them
Impact We have performed public outreach and published as a result of this collaboration.
Start Year 2016
 
Description Collaboration for surface analysis 
Organisation University of Nottingham
Department School of Pharmacy
Country United Kingdom 
Sector Academic/University 
PI Contribution With the University of Nottingham we have been able to describe the switching of the user activated surfaces using TOF-SIMS
Collaborator Contribution We supplied the materials and they supplied the expertise.
Impact The outputs have been in the form of joint publications
Start Year 2013
 
Description Collaboration with Anacail (Spin off) 
Organisation Anacail Ltd
Country United Kingdom 
Sector Private 
PI Contribution We have stabilized a collaboration with Anacail to sterilize protein coating preserving the bioactivity. This will allow the commercialization of future implants in sterilize conditions.
Collaborator Contribution The have been performing the sterilization of the samples and given ideas.
Impact We have been able to find a range of doses to preserve the bioactivity of the big protein coatings and sterilize the samples at the same time. In the future we are going to be working towards finding the right amount of doses to preserve the bioactivity of smaller molecules (such as growth factors).
Start Year 2017
 
Description Collaboration with Bone Cancer Research Trust 
Organisation Bone Cancer Research Trust
Country United Kingdom 
Sector Charity/Non Profit 
PI Contribution We are developing models of sarcoma that are useful for bone cancer research
Collaborator Contribution They are supporting us with training placements, access to patients and advice.
Impact Development of better tissue models and enhanced training.
Start Year 2020
 
Description Souhtampton Collaboration 
Organisation University of Southampton
Department Bone and Joint Research Group
Country United Kingdom 
Sector Academic/University 
PI Contribution This is a long term collaboration where cells, materials and expertise are shared to drive research
Collaborator Contribution This is a long term collaboration where cells, materials and expertise are shared to drive research
Impact A large number of papers, grants and IP.
 
Description 'I'm a ScienceGrrl, talk to me!' event at Glasgow Science Festival 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Discussion with children on this science treasure hunt sparked questions and discussion about my research area.

The children appeared to have a greater understanding of what a scientist was, and learned a little about cells and my research, and the adults accompanying the children also appeared engaged in the activity.
Year(s) Of Engagement Activity 2014
 
Description Cell block science 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Other audiences
Results and Impact Presented at prisoner outreach day at HMP Shotts
Year(s) Of Engagement Activity 2019
 
Description Centre for Doctoral Training Retreat 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Retreat to The Burn for formal PGR student training in entrepreneurship
Year(s) Of Engagement Activity 2021
 
Description Conference - Oral Presentation - BORS 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact 2016 BORS conference was organized in Glasgow, were I gave a talk with the title 'Engineering osteogenic coatings' and new collaborations were initiated between research and clinical groups afterwards.
Year(s) Of Engagement Activity 2016
 
Description Conference organisation CCE-GLORI 2019 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact The local CCE-GLORI workshop is organised by Dr Monica P. Tsimbouri every year. It is a workshop where postgraduate students present their work to their peers and supervisors and discussion then follows the talks.
This is highly beneficial to students and promotes collaboration.
Year(s) Of Engagement Activity 2019
 
Description Conference organisation-GLORI2020 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Dr Monica P. Tsimbouri organises GLORI bi-annually. This conference encourages collaboration between the basic sciences, applied sciences, engineering and clinic. This has the aim of bringing the latest ideas in basic materials research into use to deliver the next-generation of orthopaedic care. It combines expertise from orthopaedic surgeons, plastic surgeons, biologists, engineers and chemists.
The conference is free for attendance and involves oral and poster presentations from professionals, scientists, clinicians and postgraduate students. It is always well attended.
It is sponsored by industry who are also involved in the conference.
Year(s) Of Engagement Activity 2020
 
Description Debate on stem cell ethics 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Presentations and panel stimulated discussion with attendees.

There were articles written after the event by Oxbridge Biotech Roundtable and the Humanist Society, and I was contacted some time afterwards by a member of the public who was interested in stem cell therapy.
Year(s) Of Engagement Activity 2013
URL http://www.oxbridgebiotech.com/review/featured/obr-glasgow-the-ethical-issues-surrounding-stem-cell-...
 
Description EMBO Conference "The molecular and cellular basis of regeneration and tissue repair" (Oxford, UK) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact This conference has become the key international meeting for scientists investigating basic biological questions in the fields of regeneration and repair using a whole-animal 'systems' approach.

The meeting aims to bring together leading scientists who use different model organisms and will focus on different aspects of the underlying mechanisms that control the regenerative response.

The overall goal of this meeting is to significantly advance our understanding of how regeneration is coordinated in a whole-animal system so that those who work in systems that do not regenerate may develop more effective therapeutic approaches to convert healing into regeneration.
Year(s) Of Engagement Activity 2012
URL http://events.embo.org/12-regeneration/
 
Description European Society for Biomaterials Keynote (Dalby) Talk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Keynote conference talk
Year(s) Of Engagement Activity 2021
 
Description GLORI conference organisation 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Planning and Organisation of Dr Monica P Tsimbouri the scientific mentor of the Glasgow Orthopaedic Research Initiative (GLORI). GLORI has been established to encourage collaboration between the basic sciences, applied sciences, engineering and clinic. This has the aim of bringing the latest ideas in basic materials research into use to deliver the next-generation of orthopaedic care. It combines expertise from orthopaedic surgeons, biologists, engineers and chemists not only from Glasgow and Strathclyde Universities but the rest of Scotland too and also with the view to expand to the rest of the UK.
Year(s) Of Engagement Activity 2018
 
Description Healickick (EU grant) translation day 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Discussion between academics and surgeons around technologies we have developed.
Year(s) Of Engagement Activity 2021
 
Description Invited talk at British Orthopaedic Research Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Discussed research training of junior surgeons
Year(s) Of Engagement Activity 2021
 
Description Judge for Molecular, Veterinary and Life Sciences 3 Minute Thesis (3MT) 2014 heats 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Postgraduate students
Results and Impact The students' talks sparked discussion and insight into other areas of research.

I helped facilitate the event, where postgraduate students presented short 3MT versions of their theses as part of a public engagement project, and the winning students went on to present their research to additional audiences outside the University.
Year(s) Of Engagement Activity 2014
 
Description Keynote talk (Dalby) at Tissue Engineerign and Regenerative medicien International Society 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Keynote talk at a major conference
Year(s) Of Engagement Activity 2021
 
Description Media interest BBC (nanokicking) 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Talk sparked public interest

Many interested enquirers.
Year(s) Of Engagement Activity 2012
 
Description Media interest BBC (orthopaedic materials) 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Sparked public interest

Enquieries from the public
Year(s) Of Engagement Activity 2013
 
Description Media interest BBC (stem cell growth) 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Talk sparked public interest

Enquieries from the public
Year(s) Of Engagement Activity 2011
 
Description Nanotoxicology Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Invited talk for the Naotoxicology Society
Year(s) Of Engagement Activity 2016
 
Description Participation in an activity, workshop or similar - GLORI-2018 
Form Of Engagement Activity A formal working group, expert panel or dialogue
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Participate in the organization and submitted a research abstract, stimulating discussion and chaired a conference session on bone regeneration strategies.
Year(s) Of Engagement Activity 2016,2017,2018
 
Description Regeneration (art-science crossover project and event) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact This project stimulated a lot of discussion about stem cell biology and regenerative medicine from participants (including artists, musicians, scientists, members of the public) and those attending the final event. I gave lab tours and presentations to the participants, who made creative projects (such as musical compositions and poetry) for the final event, and some gave presentations about their experience of the lab, the science and their artistic response, which also sparked inter-disciplinary discussion.

After the event, many of the participants in the project were keen to continue their involvement with future Regeneration-style projects, and the Glasgow Science Festival staff were keen to hold a re-run of the event as part of the Glasgow Science Festival.
Year(s) Of Engagement Activity 2014
 
Description Royal Society of Chemistry Biomaterials Meeting 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Conference presentation
Year(s) Of Engagement Activity 2017
 
Description Scottish Metabolomics Network 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact Invited talk
Year(s) Of Engagement Activity 2016
 
Description TCES 2010 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation paper presentation
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Dr Monica P Tsimbouri. exiting discussion after talk

Interested parties continued discussion
Year(s) Of Engagement Activity 2010
 
Description TCES 2012 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation paper presentation
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Dr P Tsimbouri. talk sparked questions and discussion afterwards

Engagement with company Reps and received very good discount on equipment.
Year(s) Of Engagement Activity 2012
 
Description TCES 2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Scientific report
Year(s) Of Engagement Activity 2014
 
Description TERMIS EU 2011 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation paper presentation
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Dr Monica P Tsimbouri. Talk sparked questions and discussion afterwards

There was further discussion during a later session about my talk
Year(s) Of Engagement Activity 2011
 
Description Talk in Portugal 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Invited talk for Dalby at University of Aviero
Year(s) Of Engagement Activity 2022
 
Description UKSB 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Type Of Presentation paper presentation
Geographic Reach National
Primary Audience Postgraduate students
Results and Impact Dr M. P. Tsimbouri. Interesting discussion following presentation

Discussion lead to problem solving for a fellow scientist
Year(s) Of Engagement Activity 2010
 
Description Yorkhill Research Day 2014 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Professional Practitioners
Results and Impact Questions and discussion followed presentation. Poster award obtained.
Year(s) Of Engagement Activity 2014
 
Description seminar ntalk 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Professional Practitioners
Results and Impact Local biomaterial seminar series, and discusion followed the presentation.
Year(s) Of Engagement Activity 2016