Chiropteran (Hypsugo savii) Post-Natal Brain 2D-In Vitro Models: Primary Cell Isolation, Immortalization and...

Area of Science:

• Neuroscience
• Virology
• Cell Biology

Background:

• Bats are key reservoirs for zoonotic pathogens.
• Existing chiropteran neural *in vitro* models are limited.
• Understanding bat neurobiology is crucial for One Health initiatives.

Purpose of the Study:

• Establish and characterize novel chiropteran brain-derived 2D *in vitro* cell models.
• Develop immortalized cell lines for extended research.
• Provide a platform for studying bat neurobiology and infectious diseases.

Main Methods:

• Primary chiropteran brain cells (CpBCs) were isolated from *Hypsugo savii*.
• Immortalization was attempted using Simian virus 40 large T antigen (SV40) and human telomerase reverse transcriptase (hTERT).
• SV40-mediated electroporation successfully generated immortalized chiropteran brain cells (CiBCs).
• RNA sequencing was performed to compare transcriptional profiles of CpBCs and CiBCs.

Main Results:

• SV40 transfection yielded immortalized CiBCs; hTERT and liposome methods failed.
• CiBCs showed upregulated cell cycle and proliferation pathways compared to CpBCs.
• CiBCs displayed downregulated apoptosis, inflammation, and immune pathways.
• SV40 immortalization induced partial dedifferentiation, with neural stem cell-like and cancer-associated signatures.

Conclusions:

• The study presents the first 2D *in vitro* platform using immortalized bat brain cells.
• This novel platform facilitates research into bat neurobiology and host-pathogen interactions.
• The models are valuable for studying viral tolerance and neurotropic diseases in bats, relevant to zoonoses.