Tag Archives: ethics

Ethics – consent and medical questionairre

The ethics amendment process was not quite as quick as I had hoped with a few requests for revisions:

Revision Requests:

I did not think that I need to include formal participant consent forms since I am both participant and lead researcher. However, the ethics officer indicated that this is now a requirement.

While it has meant more work for me, I can see the value in including a formal consent form and medical questionnaire. It ensures that there is clear consent alongside identification of risks associated with the procedures. It also ensure that I formally declare my fitness (i.e. health) in undertaking the processes listed (skin biopsy and blood collection) and clearly list the ways in which risks could be mitigated (e.g. reduce likelihood of bruising, scarring etc). While I already had a good understanding of these parameters, it was useful to have a formal list and agreement. Completing these forms will also make sure I am prepared for future applications, especially involving external (or internal) participants.

I’ve done my best with the document (with review by Brad) so am hopeful for approval for a Feb start! Luckily, I don’t mind admin/paperwork and find the ethics process interesting 🙂

Revised Ethics – Blood cells for iPSCs

Due to COVID supply issues, we are still having issues sourcing key reagents etc. for the project. As an alternative iPSC protocol, we are now planning on using blood cells. The main reason is that it is a regular and active protocol in the broader lab area with clear in-house expertise. This also works better conceptually for me than harvesting cells from a skin biopsy – after all blood is strongly associated with notions of kinship . It is also nice to move into the footsteps of my dear colleague Dr Trish Adams who used iPSC technology to turn blood cells into heart cells for the project Machina Carnis.

This does entail a further ethics amendment, but since we have prior approval for skin biopsy harvest, I do not foresee any major issues. I hope to submit this before holidays – ready for the new year!

Ethics GRANTED! …but one more approval to go…

We have approval to move forward with the key aims of the project. This is great news as it means I can start working with own cells. I am still a bit precious as there are limited vials, so I will do a couple of weeks of training on HBVPs before I move on to my own cells.

While we can get started on the fibroid cell culture, Brad realised that cell immortalisation will require further Institutional Biosafety Committee (IBC) approval. This is because the process will require the uses of lentiviral vectors. As such, it is considered Notifiable Low Risk Dealing (NLRD) and the committee will need to ensure that we have the appropriate facilities and training in place to move forward.  iPSC cell reprogramming is exempt, but we still need to let the IBC know what we are doing.

The application is due tomorrow, so we had a meeting this morning to go over the protocols and identify the particular kits we are going to use. There are a range of biomedical research supply companies, but the important thing is to make sure that we use an organisation that has an Australian supplier.  hTERT and SV40 T Antigen kits are the best options for our project as they are suitable for a range of  cell types including fibroblasts.  Fischer Scientific have Alstem Immortalisation Kits available, but ABM may also be a good option. They also have a good overview of Cell Immortalisation Protocols for anyone interested in the process.

For iPSC reprogramming, we are going to use the Epi5™ Episomal iPSC Reprogramming Kit by Thermo Fischer. Another group has used this product previously – so we can get tips on how to get the best results.  Lovely Jo-Maree is looking into the best purchasing options. With lead time for purchase and delivery, the products will likely arrive around the same time as final approval.

Ethics update

We’ve received our first comment from the HREC Chair regarding our ethics application. They raised an important consideration in relation to more clearly outlining some of the ethical implications and questions that the project raises.  Here is my current thinking:

Posthuman Genetic Legacies raises some ethical issues in relation to the ownership and governance of biological materials. The ethical implications of using human biomaterials for scientific and artistic research form part of the investigation and will enable the research team to identify and consider these issues from creative, legal, and scientific perspectives. This initial enquiry will also form the basis for subsequent exploration which will focus more specifically on bioethics with attention to legal and ethical frameworks for the management of use of biological materials.

Some of the key issues and questions include:

  • Ownership and use of biomaterials (cells and tissue) when removed from the body including tensions arising between individual donor, research team and university.
    • Who legally owns biomaterials (cells and tissue) when they are removed from the body?
    • What rights does the original donor (and researcher-participant) legally maintain when working in a team research environment at a university?
    • What current legal and governance frameworks are in place and which aspects may need reconsideration to better accommodate the interests of all stakeholders?
  • IP implications to produce biomaterials in a university environment
    • How is IP negotiated within a university environment when biomaterials are associated directly with an individual researcher as participant?

Once primary cells are successfully immortalised, the research team will  review current policies regarding the use of biomaterials for artistic use in  Australia. To date, most literature on biological art focuses on the conceptual, ethical and theoretical affordances of the practice and basic lab protocols with limited insight into legal and  governance frameworks, especially where commercial and research interests intersect.

While the creation of the Billy Apple® cell line was a success (Hilton 2014), claims regarding the uptake of the cell line as part of the ATCC are difficult to verify as the cell line is currently not listed in the online cell product listing. Creating another artist cell line for uptake into the American Type Culture Collection (ATCC) or alternative distributor will enable researchers to gain insight into the current policies and practices that underpin biomaterials use, storage and distribution. These insights can then be used to compare policy documentation and experiences of research from art and science over the past 20 years. This will occur in the next stage of project development.

During this phase, the research team will also consider how ethical implications shift when moving from a university research setting to a more open/shared or commercial research environment such as the storage and distribution of biomaterials by companies such as the ATCC.  During this stage, questions will address implications for biomaterial distribution and interdisciplinary engagement:

Some of the key issues and questions include:

  • Ownership and use of biomaterials including tensions arising between individual donor, family, research team, universities, and external organisations such as biomaterial distribution companies.
    • Who legally owns biomaterials (cells and tissue) when they become part of a global biomaterials repository?
    • What ethical guidelines currently govern the use of biomaterials in a global setting? Can you and should you control or restrict the use of biomaterials in relation to personal or cultural values?
    • Should extended family members have input into the distribution of biomaterials (and associated personal information)?
  • IP implications to produce biomaterials across disciplinary terrains (who owns IP when research and commercial interests are involved)
    • How is IP negotiated within a university environment and commercial research setting when biomaterials are associated directly with an individual researcher as participant?
  • Privacy issues for disclosure of personal information (personal details and genetic information) in relation to biomaterials use and distribution.
    • What are the potential risks (short term and long term) involved in the disclosure of personal and genetic information in relation to biomaterials in a research setting (used ​and stored within universities) versus commercial research context (stored and distributed by an external company)?
    • What biomaterial information is useful for scientific versus artistic disciplines?
    • What processes should be put in place to protect the privacy of the donor but still provide useful information across disciplinary domains?
  • Communication including public understanding and transparency of regulatory and governance frameworks in biomaterials research across different domains?
    • What is the public perception and research value of establishing formal frameworks for the development of biobanks and biomaterials for use in artistic research?

This stage will involve further ethical clearance and require me to work with the research team to develop a cohesive research plan and identify ways of minimising risk in terms of privacy.

While I am somewhat familiar with the ethics clearance process, I am new to thinking through the implications as participant where personal medical and genetic information and biomaterial may become shared research materials. This is where collaboration with experts from the Centre for Law and Genetics is vitally important. I look forward to hearing from my collaborator Jane (and hope I was not completely off track).

Hilton, C., 2014. The immortalisation of Billy Apple®: an art-science collaboration. Leonardo47(2), pp.109-113.

Ethics in!

Brad has sent through final information for Stage 2 ethics clearance.  We included a skin biopsy protocol in case the fibroid cells are unviable. Unfortunately, when dealing with primary cells, nothing is certain and there is a possibility that the initial cells are contaminated or do not recover from the freezing process. As such, we have included ‘Plan B’ to ensure we can move forward with the project and establish a new batch of primary cells for immortalisation.

The ethics documents were formally submitted through the Ethics Review Manager. Hopefully, I have addressed all project aspects and provided sufficient information for approval.  Fingers crossed!

Project Meeting

We had a detailed project planning and ethics clearance review meeting on Monday. It was heartening to hear that the ethics clearance documentation is almost ready for formal submission.

These are some of the processes we are expecting to undertake:

  • Cell culture of fibroid cells (optimisation of culture methods for 2D and 3D environments, cellular response and proliferation testing)
  • Co-culture of fibroid cells with other cell lines
  • Genetic profiling of primary fibroid cells
  • Immortalisation of primary fibroid cells via established commercial kit (e.g. Applied Biological Materials (ABM) cell immortalisation kits)
  • Reprogramming of primary cells to generate induced pluripotent stem cells (iPSCs) via established commercial kit (e.g. Epi5™ Episomal iPSC Reprogramming Kit available via Thermo Fisher)
  • Cell culture of reprogrammed or immortalised fibroid cells (optimisation of culture methods for 2D and 3D environments, cellular response and proliferation testing)
  • Genetic profiling of reprogrammed or immortalised fibroid cells and cells lines (if successful)
  • Fixing and staining of cells
  • Light and confocal microscopy of cultured cells
  • Timelapse video of cultured cells

Other potential processes include:

  • Green Fluorescent Protein (GFP) cell tagging [UTAS]
  • Scanning Electron Microscopy (SEM) of cultured cells [UTAS]
  • Transmission Electron Microscopy (TEM) of cultured cells [UTAS]
  • Histopathology of cultured 3D structures [UTAS or QUT]
  • Development of Gastruloids, Organoids or Neurospheres (self-organised 3D cell masses) [UTAS]

I was delighted to hear that my desires for creating aggregates of cells via the production of gastruloids or organoids was not outside the domain of possibility. Jo-Maree has produced neurospheres (balls of neural stem cells) previously, so there may be scope (and hopefully time) to experiment with cell clusters.

Gastuloids are of particular interest to me as they are cell clusters that display features of early embryo development. Staining and fluorescence imaging (apart from being visually stunning) enables the visualisation of tissue organisation as shown in this figure and caption from the Nature publication ‘Multi-axial self-organization properties of mouse embryonic stem cells into gastruloids’:

Tissue organisation in gastruloids

Tissue organization in gastruloids a, Gastruloids formed from Sox1GFP;BramCherry (SBR) line and stained for Sox2 expression (Sox1GFP and SOX2 signals are displayed in green and magenta, respectively). White arrowheads indicate tubular SOX2/Sox1-positive neural structures. Red arrowheads point to the presumptive digestive tube. b, WISH on 8-µm transverse cryosections of gastruloids at 144 h AA using Sox2 and Meox1 antisense probes, counter-stained with Nuclear Fast Red. Sox2-positive cells localized predominantly in a compact dorsal domain, whereas Meox1 signals were found in two bilateral domains. The domain of expression of each gene is outlined with white dashed lines. c, Haematoxylin and eosin staining of transverse paraffin sections of different gastruloids at 120 h AA, showing the diversity of cell types and several levels of tissue organization. d, Gastruloids formed from Sox1GFP;BramCherry ESCs were fixed and stained at 168 h AA for OLIG2 (top, white), PAX3 (middle, red) and PAX7 (bottom, red). Scale bars as indicated. c, d, Gastruloids formed from Sox1GFP; BramCherry ESCs collected at 168 h AA and stained for SOX17 (magenta, c) or CDX2 (magenta, d). Scale bars as indicated.

Gastruloids are created from embryonic stem cells, although iPSC cells (cells that have been reprogrammed into a stem cell like state) have also been used. As such, if I have success with reprogramming my fibroid cells to iPSC cells, I could use them to make, and learn more about, gastruloids.

During our discussion, we decided it would be a good idea to include an optional alternative to the fibroid cells – just in case there is an issue with contamination or the freezing/thawing process. While I have isolated skin cells (fibroblasts) from hair follicles and skin grafts previously as part of the HSE Project at QUT, we decided on skin scrapings.   This approach was selected as will enable the isolation of skin cells, is not too invasive and is well established within the School of Medicine.

Once I’ve included this information including the protocol, I should be ready to submit the final ethics clearance document.

Beccari, L., Moris, N., Girgin, M., Turner, D.A., Baillie-Johnson, P., Cossy, A.C., Lutolf, M.P., Duboule, D. and Arias, A.M., 2018. Multi-axial self-organization properties of mouse embryonic stem cells into gastruloids. Nature562(7726), pp.272-276.

Next Step: Ethics Clearance

I received formal ethics clearance in 2020 for the isolation of primary cells from my fibroid tissue.  One of the next key elements to move the project forward is to receive ethical clearance (via amendment to the original application) for key laboratory processes including cell immortalisation and induced pluripotent stem cells (iPSC).

When I was a PhD student, I initially viewed the process as an awkward and time-consuming hurdle. I thought it was mainly to protect the institution from liability. However, as I have learned more about the ethics review process – I am now even a new member of the UTAS Human Research Ethics Committee HREC –  I recognise the importance of ethics clearance for ensuring the safety of researcher and participants. As part of the ethics clearance process, I am required to put together a project overview. This covers research team details, research questions and aims, supporting literature, project design (methodology and methods), data collection and management and future directions. While it does take time to compile the information, it is a useful tool to think through project parameters clearly and articulate the value of the project beyond my own interests. It also ensures that there are clear guidelines for protecting and working with participant data. While, my project is only concerned with my own biomaterials and data, it is still important to set up a storage and backup system with password protected files for team and private (e.g. personal medical details) information.

I have completed my amendment and am waiting for Brad and Jo-Maree’s input before formally submitting the document for review by the HREC. I may have been a bit ambitious in my scope of what may be possible to do as part of an eight-month residency. For example, I am keen to try and establish gastruloids or organoids, but this may be too ambitious as it would rely on successful iPSC. This is where team input is vital to ensure feasibility.