Poster Presentation 37th Lorne Cancer Conference 2025

Establishment characterisation and utilisation of nine novel patient derived osteosarcoma cell lines (#106)

Zara A Barger 1 2 , Ashleigh M Fordham 1 2 3 , Jie Mao 1 2 , Lauren Brown 1 2 3 , Callum M Perkins 1 2 , Alice Salib 1 2 3 , Kate Gunther 1 2 , Paul G Ekert 1 2 3 4 5 , Emmy DG Fleuren 1 2 3
  1. UNSW Medicine and Health, UNSW Sydney, School of Clinical Medicine, Sydney, NSW, Australia
  2. Children's Cancer Institute, Kensington, NSW, Australia
  3. University of New South Wales Centre for Childhood Cancer Research, UNSW Sydney, Sydney, NSW, Australia
  4. Murdoch Children's Research Institute, Melbourne, VIC, Australia
  5. Peter MacCallum Cancer Centre , Melbourne, VIC, Australia

Background

Paediatric osteosarcoma survival rates remain largely unimproved over the past three decades, with only a 20% five year survival rate for recurrent or metastatic disease. The highly heterogeneous nature of osteosarcoma requires the development of robust in vitro models for the pre-clinical testing of novel therapeutics, which is critical for identifying more effective and less toxic treatments. Previous attempts at developing patient-derived osteosarcoma cell cultures have had limited success. Here we describe our program to generate new osteosarcoma cell lines derived from patient samples, expanding the resources available for research.

Methods

Patient-derived xenograft (PDX) tumours were harvested, dissociated, and depleted of mouse cells. The tumour cell populations were cultured in multiple conditions to determine optimal culture medium, supplementation, and plate-coating to maximise viability and growth rate. Culture origin was validated by comparing copy-number profiles, determined by SNP array, to whole genome sequencing of original patient tumours. Immunohistochemistry (IHC) was used to verify tumour content. Cell lines were also transduced with the CRISPR-Cas9 system and subjected to short- and long-term drug viability assays, to determine potential gene editing and therapeutic applications.

Results

We identified a protocol that allowed in vitro osteosarcoma cell expansion while retaining the patients’ original tumour characteristics. Utilising this protocol, we successfully established nine validated, unique patient-derived osteosarcoma cell lines (OS-PDX) from a total of 13 successfully engrafted osteosarcoma PDXs. We successfully transduced OS-PDX cell lines with CRISPR-cas9, and revealed known and novel drug dependencies, illustrating their research potential.

Discussion/Conclusion

The establishment of robust osteosarcoma models significantly extends resources available that allow preclinical exploration of osteosarcoma biology and new targeted therapies. Early data illustrates their research potential, including identifying potential novel therapeutic avenues, which we are currently exploring in matched osteosarcoma in vivo models.