Optimising an Experimental In Vitro Blood-Brain Barrier Model for Cancer Migration Studies
Author
Pyziak, Sonia Agnieszka
Term
4. term
Education
Publication year
2026
Submitted on
2026-05-28
Pages
63
Abstract
Breast cancer and melanoma often spread to the brain. When tumor cells get past the blood–brain barrier (BBB)—the brain’s protective border—the disease becomes more aggressive and harder to treat. The BBB tightly controls what moves from the bloodstream into brain tissue, which is essential for normal brain function but also makes it difficult to deliver treatments for brain diseases, including brain metastases. This project set out to build a simplified laboratory version of the BBB that can be used to study how cancer cells move across it. The model was created in Transwell co-culture inserts with either 1 μm or 8 μm pores and was lined with primary brain microvascular endothelial cells (BMECs) freshly isolated from mouse brains. We carried out extensive optimization to improve the model’s stability and the formation of tight junctions—the protein seals between endothelial cells that give the BBB its selectivity. We confirmed barrier formation by measuring high transendothelial electrical resistance (TEER), a standard readout of how “tight” the barrier is. To test whether cancer cells could cross, we labeled breast (4T1 em GFP) and melanoma (B16F10 e-GFP) cell lines with green fluorescent protein (GFP) so they could be tracked. Their movement suggested that passage through the Transwell membranes depended on pore size. The BBB model is still exploratory and will need further refinement to improve reproducibility and make it more relevant to human disease. Even so, these results support further development of the model as a platform for studying cancer cell migration and for identifying ways to limit the spread of brain metastases.
Brystkræft og melanom spreder sig ofte til hjernen. Når kræftceller passerer blod-hjerne-barrieren (BBB) – hjernens beskyttende grænse mod blodet – bliver sygdommen mere aggressiv og sværere at behandle. BBB styrer stramt, hvad der kan passere fra blodet ind i hjernevævet. Det er afgørende for normal hjernefunktion, men gør det også svært at levere behandlinger mod hjernesygdomme, herunder hjernemetastaser. Formålet med dette projekt var at opbygge en forenklet laboratoriemodel af BBB, der kan bruges til at undersøge, hvordan kræftceller bevæger sig over barrieren. Modellen blev lavet i Transwell-co-kultursystemer med enten 1 μm eller 8 μm porer og blev beklædt med primære hjernemikrovaskulære endotelceller (BMECs) frisk isoleret fra musehjerner. Vi gennemførte omfattende optimeringer for at øge modellens stabilitet og dannelsen af tight junctions – de proteintætninger mellem endotelceller, som giver BBB sin selektivitet. Vi bekræftede barrieredannelsen ved at måle høj transendothelial electrical resistance (TEER), en standardmåling for, hvor "tæt" barrieren er. For at teste om kræftceller kunne krydse, mærkede vi brystkræft- (4T1 em GFP) og melanom- (B16F10 e-GFP) cellelinjer med grønt fluorescerende protein (GFP), så de kunne spores. Deres bevægelse tydede på, at passagen gennem Transwell-membranerne var afhængig af porestørrelsen. BBB-modellen er stadig på et indledende stadie og kræver yderligere forbedringer for at øge reproducerbarheden og den kliniske relevans. Alligevel peger resultaterne på, at modellen kan videreudvikles som platform til at studere kræftcellers migration og til at identificere måder at begrænse spredningen af hjernemetastaser.
[This apstract has been rewritten with the help of AI based on the project's original abstract]
Keywords