# Establishment of an Immortalized Canine Hippocampal Neural Stem Cell Line via SV40LT Retroviral Transduction

**Authors:** Yankun Ke, Zixin Li, Huaiyu Wang, Yixuan Zhang, Shiyu Xu, Longlong Zhang

PMC · DOI: 10.3390/cells15060543 · 2026-03-19

## TL;DR

Scientists created a long-lasting dog neural stem cell line that can be used to study brain diseases and potential treatments.

## Contribution

The novel contribution is the successful establishment of an immortalized canine hippocampal neural stem cell line using SV40LT retroviral transduction.

## Key findings

- The immortalized NSC line can be passaged indefinitely while maintaining a normal karyotype.
- Both immortalized and normal NSCs differentiate into neuronal and glial lineages under induced conditions.
- The line provides a valuable tool for studying neural differentiation and neurological disorders in dogs.

## Abstract

Dogs represent a promising animal model for analyzing human neurodegenerative diseases, owing to their similarities to humans in nervous system architecture and behavioral phenotypes. Neural stem cells (NSCs) serve as a highly valuable in vitro experimental model for investigating neurogenesis, neurodegenerative disease pathogenesis, and neural molecular biology; however, studies on immortalized canine neural stem cell lines remain scarce. Herein, we successfully established an immortalized canine hippocampal neural stem cell line that can be continuously passaged in vitro via SV40 large T antigen (SV40LT) viral infection and subsequent cellular transformation. Both the immortalized NSCs and their normal parental counterparts differentiated into neuronal and glial lineages under induced differentiation conditions. Normal canine hippocampal NSCs can be passaged for no more than 10 generations, whereas the immortalized line can be passaged indefinitely while maintaining a normal karyotype. This immortalized canine hippocampal NSC line can act as a critical experimental tool for future research into neural differentiation mechanisms and stem cell-derived therapeutic strategies for neurological disorders in dogs.

## Linked entities

- **Species:** Canis lupus familiaris (taxon 9615)

## Full-text entities

- **Genes:** PDGFA (platelet derived growth factor subunit A) [NCBI Gene 491597], Nes (nestin) [NCBI Gene 18008] {aka ESTM46, Ifaprc2, Marc2, RC2}, Pdgfra (platelet derived growth factor receptor, alpha polypeptide) [NCBI Gene 18595] {aka CD140a, Pdgfr-2}, Caps2 (calcyphosphine 2) [NCBI Gene 353025] {aka D630005B03Rik}, SOX2 (SRY-box transcription factor 2) [NCBI Gene 488092], GDNF [NCBI Gene 489224], Mki67 (antigen identified by monoclonal antibody Ki 67) [NCBI Gene 17345] {aka D630048A14Rik, Ki-67, Ki67}, VIM (vimentin) [NCBI Gene 477991], Map2 (microtubule-associated protein 2) [NCBI Gene 17756] {aka G1-397-34, MAP-2, Mtap-2, Mtap2, repro4}, EGF (epidermal growth factor) [NCBI Gene 403657] {aka CEGF}, LOC106557476 (tubulin alpha-1A chain) [NCBI Gene 106557476] {aka TUBA1A}, BDNF (brain derived neurotrophic factor) [NCBI Gene 403461], FGF2 (fibroblast growth factor 2) [NCBI Gene 403857] {aka BFGF}, Gfap (glial fibrillary acidic protein) [NCBI Gene 14580], CADPS2 (calcium dependent secretion activator 2) [NCBI Gene 475304], GFAP (glial fibrillary acidic protein) [NCBI Gene 480495], Syn1 (synapsin I) [NCBI Gene 20964] {aka Syn-1, Syn1-S}, Sox2 (SRY (sex determining region Y)-box 2) [NCBI Gene 20674] {aka Sox-2, lcc, ysb}
- **Diseases:** CNSC (MESH:D004283), neurological disorders (MESH:D009461), Alzheimer's disease (MESH:D000544), necrosis (MESH:D009336), aggression (MESH:D010554), viral infection (MESH:D014777), neurodegenerative disease (MESH:D019636), neurological and psychiatric diseases (MESH:D001523), neuroinflammation (MESH:D000090862), cognitive decline (MESH:D003072), NSC-Ts (OMIM:617394), Retroviral Infection (MESH:D000071297), injury to (MESH:D014947)
- **Chemicals:** water (MESH:D014867), polyacrylamide (MESH:C016679), TO (MESH:D014425), puromycin (MESH:D011691), N2 (MESH:D009584), Giemsa (MESH:D001399), Colcemid (MESH:D003703), ascorbic acid (MESH:D001205), hematoxylin (MESH:D006416), DMSO (MESH:D004121), poly-L-ornithine (MESH:C008973), penicillin (MESH:D010406), methanol (MESH:D000432), Amino Acids (MESH:D000596), db-cAMP (MESH:D003994), Formalin (MESH:D005557), paraffin (MESH:D010232), Glutamax (MESH:C054122), propidium iodide (MESH:D011419), sodium citrate (MESH:D000077559), paraformaldehyde (MESH:C003043), 4',6-diamidino-2-phenylindole (MESH:C007293), SDS (MESH:D012967), acetic acid (MESH:D019342), Triton X-100 (MESH:D017830), ethanol (MESH:D000431), KCl (MESH:D011189), 3,3'-diaminobenzidine (MESH:D015100), streptomycin (MESH:D013307), polybrene (MESH:D006583), DMEM-F12 (-)
- **Species:** Canis lupus familiaris (dog, subspecies) [taxon 9615], Homo sapiens (human, species) [taxon 9606], Mus musculus (house mouse, species) [taxon 10090]
- **Cell lines:** HEK293T — Homo sapiens (Human), Transformed cell line (CVCL_0063), CNSC-T — Homo sapiens (Human), Human papillomavirus-related endocervical adenocarcinoma, Cancer cell line (CVCL_1U15)

## Figures

7 figures with captions in the complete paper: https://tomesphere.com/paper/PMC13025826/full.md

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Source: https://tomesphere.com/paper/PMC13025826