Presentations

Jeanne Loring “Stem Cells: How it Started and Where it’s Going”

Professor Emeritus at Scripps Research Institute, Jeanne Loring opened the workshop on September 17th. Jeanne recounted her journey in stem cell biology for conservation, presenting on the generation of the first iPSCs from endangered species. As a poignant link to our workshop, Jeanne brought us back to that pivotal first meeting in Vienna in 2015 where the plan for using iPSCs to save the Northern White Rhino was hatched. Finally, a reminder to test for genetic stability in iPSC lines set the tone for a scientifically rigorous meeting. 

“Don’t be afraid to understand your cells in every possible way” – Jeanne Loring

 

Keynote Speakers: Katsuhiko Hayashi and Martin Pera

Workshop participants had the privilege of hearing from two seminal pioneers in stem cell biology. 

On Day 2, Professor Katsuhiko Hayashi of Osaka University delivered a remarkable presentation exploring his cutting-edge research into in vitro gametogenesis and provided a timeline for the generation of gametes for select endangered species.

To kick off the third and final day, stem cell pioneer Professor Martin Pera of The Jackson Laboratory provided his perspectives on the spectra of pluripotency and the feasibility of using iPSC technology within conservation.  

 

Advances & Innovations

Participants enjoyed four carefully selected talks designed to communicate out-of-the-box innovations in stem cell research. Over one hour, the audience was exposed to thought-provoking stem cell research that touched on artificial intelligence, high-throughput differentiation to any cell type, coral conservation, and interspecies chimeras.

• Stuart Chambers, Brightfield Therapeutics
• Alex Ng, GC Therapeutics
• Nikki Traylor-Knowles, University of Miami
• Jun Wu, University of Texas Southwestern

 

Lightning Talks

Showcasing some of the incredible work performed by the people in the room, a rapid-fire series of lightning talks invigorated and inspired. We heard about endangered felid reprogramming in Thailand, the use of micro-gels to transform cell culture, and creating a village-in-a-dish using wolf iPSCs. Topics spanned de-extinction, reptilian embryogenesis, and industry perspectives, in addition to biobanking in Singapore and, of course, cloning.

Proof-of-principle for pooled biopreservation using gray wolf (Canis lupus) samples: a village-in-a-capillary approach

Francisco Pelegri University of Wisconsin-Madison 

 

Improved reprogramming efficiency using species-specific factors

Sara Ord Colossal Biosciences 

 

The Diversity of Squamate Pre-Oviposition Embryogenesis

Antonia Weberling University of Oxford 

 

Challenges to generate non-integrative felid iPSCs

Woranop Sukparangsi Burapha University 

 

Creating solutions and services for somatic reprogramming

Uma Lakshmipathy Thermo Fisher Scientific 

 

Wider technological crossroads for Animal Cloning and Stem Cells and their opportunities for conservation 

Andrew French Cartherics 

 

Endangered Species Conservation via Assisted Reproduction

Nicole Tay Institute of Molecular and Cell Biology 

 

Cells in gels – promoting pluripotency in 3D microgels

Timo Kohler University of Cambridge 

 

Discussion Panels

A significant portion of the meeting was devoted to transparent discussion. To accelerate the sharing of information and promote active audience engagement, the meeting included four Panel-Led Discussion Sessions.

 

Reprogramming: Progress and Pitfalls

First reported in 2006 by Yamanaka and Takahashi, induced pluripotent stem cells or ‘iPSCs’ can be produced from any cell in the body, in a process known as ‘reprogramming’. These remarkable cells mirror the earliest stage of embryonic development and can become any cell or tissue type, including gametes (sperm and eggs). This capacity makes them tremendously useful for biobanking and has the power to level the playing field within wildlife conservation. 

The biggest challenge is getting this process to work across the planet’s highly diverged taxa. There are many different methods for inducing pluripotency, and these are often shown to be species-specific. An intensive discussion surrounding reprogramming was led by a unique panel spanning expertise from a broad range of species. 

• Manabu Onuma, National Institute for Environmental Studies
• Deanne Whitworth, University of Queensland
• Jun Wu, University of Texas Southwestern
• Thomas Zwaka, Icahn School of Medicine at Mount Sinai 

Our experts discussed what has worked best for each of their model species and speculated on the importance of reproducing ‘naive’, or, ‘ground-state’ pluripotency. Deanne Whitworth stimulated discussion around the most recent data for pluripotent states, while Thomas Zwaka and Jun Wu spoke about their research deriving iPSCs from a range of mammalian species. Representing the non-mammalian experts, Manabu Onuma outlined how to make iPSCs from endangered Japanese birds.

The general consensus was that we are currently too dependent on data from mouse and human stem cells. Reprogramming in a new species often requires years of optimization, and pluripotency for non-model species is not well understood. Nevertheless, the room converged around positivity for the feasibility of this technology across species. In the audience, Oliver Ryder of the San Diego Zoo Wildlife Alliance spoke about the importance of advancing our understanding and incorporating these tools in conservation efforts.

 

Exploring Methods for In Vitro Gametogenesis 

One of the most promising applications of iPSC technology for conservation is the generation of sperm and eggs from these cells. This process is known as in vitro gametogenesis and has been effectively demonstrated in rodent models leading to viable offspring with the capacity to breed. 

To explore the topic, we assembled participants with a range of expertise, who use a variety of model species and different methods to derive germ cells in culture. 

• Ruth Appletant, University of Antwerp
• Qilong Ying, University of Southern California
• Katsuhiko Hayashi, Osaka University
• Marisa Korody, San Diego Zoo Wildlife Alliance

The panel discussed alternative approaches to deriving germ cells and provided their perspectives on choosing a model species to advance the science. We heard from Marisa Korody about the progress and challenges her team has experienced as they try to produce primordial germ cells for the rhino. Katsuhiko Hayashi, our Keynote speaker, answered some of the audience’s most pressing questions regarding the most advanced IVG protocols we have in the world. Qilong Ying spoke to the potential use of avian models to accelerate this research, and Ruth Appeltant reminded us that we should be learning from the endogenous stem cell populations within the ovary.

The room converged on the profound promise of this technology. In vitro gametogenesis methods work well for rodent models, with promising data coming through for rhinos and avian species. As a recurring theme, the consensus was that we urgently need to develop this technology for other non-model species. Robust protocols for IVG in endangered species will transform our ability to rescue those on the brink who desperately need prioritization. 

 

Disease Modeling for Species Recovery

Wildlife disease is a major problem today, not only for endangered species but also for human health. Stem cells have become essential tools for the study of human disease and should be used in the same way to understand, predict, and treat wildlife diseases. We assembled a formidable panel to provoke discussion in this space.

• Benyamin Rosental, Ben Gurion University
• Tom Burdon, The Roslin Institute
• Gareth Sullivan, Occam BioSciences
• Andrea Bodnar, Gloucester Marine Genomics Institute

During engagement with the audience, the panel suggested a pipeline to eradicate key wildlife diseases. This involved iPSC differentiation in-a-dish to study the effects of viruses, and could ultimately result in vaccine discovery. Gareth Sullivan of Oslo University suggested ways to use organoids derived from iPSCs in disease research, while Andrea Bodnar highlighted the fascinating way her team uses stem cells from marine invertebrates in longevity research. All the way from the Roslin Institute, Tom Burdon spoke about his research using immune cells derived from pig iPSCs to study resistance to African Swine Fever. The audience was lucky to hear from Benyamin Rosental, who is taking his immunology expertise in human health to ground-breaking conservation of corals.

The group lamented that we are always in crisis mode, reacting to diseases too late. We need better ways to predict and detect disease. All spoke to the importance of developing this technology across a range of species and reminded the audience that reprogramming and differentiation protocols need to come first. 

 

Biobanking stem cells: obstacles and best practices 

This final discussion session was deliberately placed after participants had the chance to learn and engage with the latest tools in stem cell technology. Biobanks all over the world are working to preserve tissue, cell lines, and gametes from as many species as possible. Stem cells are the fundamental missing piece of the puzzle, transforming a limited resource into an unlimited one. While this potential is beginning to be recognized, there are significant challenges inherent in bringing reprogramming to applied biobanking. We heard from some of the most important figures working tirelessly to take biobanking to the next level.

• Oliver Ryder, San Diego Zoo Wildlife Alliance
• Pierre Comizzoli, Smithsonian Conservation Biology Institute
• Suzannah Williams, Nature’s Safe
• Linda Penfold, South-East Zoo Alliance For Reproduction and Conservation

Our panel had many insightful comments and perspectives on this complex and critical topic. Suzannah Williams, Pierre Comizzoli, and Linda Penfold spoke about their passion for the work and the essential inclusion of iPSC technology, and in vitro gametogenesis in biobanking. The group agreed that stem cell technology will not replace standard cell culture, but rather augment current approaches, providing an unlimited alternative cell population that can be used to make gametes, maybe even embryos, and study critical wildlife diseases. 

Oliver Ryder had the last word, imploring that we consider how we want to approach the next 1000 years. He firmly asserted that we have a moral imperative to bank the biodiversity of our planet for our future scientists. 

 

Taking The Next Step

As a follow-up to our workshop, Revive & Restore is now more interested than ever in developing a research program to fund innovative projects that advance stem cell technology for use in the genetic rescue of wildlife. We scratched the surface of this challenge during our intense brainstorming sessions at the workshop. The next step is to create a robust roadmap charting potential avenues toward improved stem cell capabilities for wild species. Having such a plan is a prerequisite to attracting serious interest from potential financial supporters, which would, in turn, enable us to issue a broad call for proposals. Our Biotech For Birds Program had similar beginnings and is now funding eight international teams to develop cutting-edge technologies for the genetic rescue of avian species.