Limits...
A Triple Culture Model of the Blood-Brain Barrier Using Porcine Brain Endothelial cells, Astrocytes and Pericytes.

Thomsen LB, Burkhart A, Moos T - PLoS ONE (2015)

Bottom Line: The BECs retain their expression of tight junction proteins and efflux transporters leading to high trans-endothelial electric resistance (TEER) and low passive paracellular permeability.The BECs, astrocytes and pericytes are often isolated from small rodents.Larger species as cows and pigs however, reveal a higher yield, are readily available and have a closer resemblance to humans, which make them favorable high-throughput sources for cellular isolation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Neurobiology, Department of Health Science and Technology, Aalborg University, Aalborg, Denmark.

ABSTRACT
In vitro blood-brain barrier (BBB) models based on primary brain endothelial cells (BECs) cultured as monoculture or in co-culture with primary astrocytes and pericytes are useful for studying many properties of the BBB. The BECs retain their expression of tight junction proteins and efflux transporters leading to high trans-endothelial electric resistance (TEER) and low passive paracellular permeability. The BECs, astrocytes and pericytes are often isolated from small rodents. Larger species as cows and pigs however, reveal a higher yield, are readily available and have a closer resemblance to humans, which make them favorable high-throughput sources for cellular isolation. The aim of the present study has been to determine if the preferable combination of purely porcine cells isolated from the 6 months old domestic pigs, i.e. porcine brain endothelial cells (PBECs) in co-culture with porcine astrocytes and pericytes, would compare with PBECs co-cultured with astrocytes and pericytes isolated from newborn rats with respect to TEER value and low passive permeability. The astrocytes and pericytes were grown both as contact and non-contact co-cultures as well as in triple culture to examine their effects on the PBECs for barrier formation as revealed by TEER, passive permeability, and expression patterns of tight junction proteins, efflux transporters and the transferrin receptor. This syngenic porcine in vitro BBB model is comparable to triple cultures using PBECs, rat astrocytes and rat pericytes with respect to TEER formation, low passive permeability, and expression of hallmark proteins signifying the brain endothelium (tight junction proteins claudin 5 and occludin, the efflux transporters P-glycoprotein (PgP) and breast cancer related protein (BCRP), and the transferrin receptor).

No MeSH data available.


Related in: MedlinePlus

Mannitol permeability measurements on PBECs in thirteen co-culture combinations as a function of their TEER.TEER measured just before the permeability experiment was conducted. The Papp mannitol measured on n = 3 for each culture condition. Each point represents one hanging culture insert with PBECs.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4524625&req=5

pone.0134765.g003: Mannitol permeability measurements on PBECs in thirteen co-culture combinations as a function of their TEER.TEER measured just before the permeability experiment was conducted. The Papp mannitol measured on n = 3 for each culture condition. Each point represents one hanging culture insert with PBECs.

Mentions: The optimal properties of an in vitro BBB model are reflected in high expression of tight junction proteins that do not just lead to a high TEER but also low passive permeability of low-molecular substances like sodium fluorescein or mannitol from the luminal to the abluminal side of the in vitro BBB model. The permeability to mannitol was measured in cultures of PBECs in thirteen different culture conditions and plotted against TEER values measured on the same PBECs just before the permeability experiments was initiated (Fig 3).


A Triple Culture Model of the Blood-Brain Barrier Using Porcine Brain Endothelial cells, Astrocytes and Pericytes.

Thomsen LB, Burkhart A, Moos T - PLoS ONE (2015)

Mannitol permeability measurements on PBECs in thirteen co-culture combinations as a function of their TEER.TEER measured just before the permeability experiment was conducted. The Papp mannitol measured on n = 3 for each culture condition. Each point represents one hanging culture insert with PBECs.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4524625&req=5

pone.0134765.g003: Mannitol permeability measurements on PBECs in thirteen co-culture combinations as a function of their TEER.TEER measured just before the permeability experiment was conducted. The Papp mannitol measured on n = 3 for each culture condition. Each point represents one hanging culture insert with PBECs.
Mentions: The optimal properties of an in vitro BBB model are reflected in high expression of tight junction proteins that do not just lead to a high TEER but also low passive permeability of low-molecular substances like sodium fluorescein or mannitol from the luminal to the abluminal side of the in vitro BBB model. The permeability to mannitol was measured in cultures of PBECs in thirteen different culture conditions and plotted against TEER values measured on the same PBECs just before the permeability experiments was initiated (Fig 3).

Bottom Line: The BECs retain their expression of tight junction proteins and efflux transporters leading to high trans-endothelial electric resistance (TEER) and low passive paracellular permeability.The BECs, astrocytes and pericytes are often isolated from small rodents.Larger species as cows and pigs however, reveal a higher yield, are readily available and have a closer resemblance to humans, which make them favorable high-throughput sources for cellular isolation.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Neurobiology, Department of Health Science and Technology, Aalborg University, Aalborg, Denmark.

ABSTRACT
In vitro blood-brain barrier (BBB) models based on primary brain endothelial cells (BECs) cultured as monoculture or in co-culture with primary astrocytes and pericytes are useful for studying many properties of the BBB. The BECs retain their expression of tight junction proteins and efflux transporters leading to high trans-endothelial electric resistance (TEER) and low passive paracellular permeability. The BECs, astrocytes and pericytes are often isolated from small rodents. Larger species as cows and pigs however, reveal a higher yield, are readily available and have a closer resemblance to humans, which make them favorable high-throughput sources for cellular isolation. The aim of the present study has been to determine if the preferable combination of purely porcine cells isolated from the 6 months old domestic pigs, i.e. porcine brain endothelial cells (PBECs) in co-culture with porcine astrocytes and pericytes, would compare with PBECs co-cultured with astrocytes and pericytes isolated from newborn rats with respect to TEER value and low passive permeability. The astrocytes and pericytes were grown both as contact and non-contact co-cultures as well as in triple culture to examine their effects on the PBECs for barrier formation as revealed by TEER, passive permeability, and expression patterns of tight junction proteins, efflux transporters and the transferrin receptor. This syngenic porcine in vitro BBB model is comparable to triple cultures using PBECs, rat astrocytes and rat pericytes with respect to TEER formation, low passive permeability, and expression of hallmark proteins signifying the brain endothelium (tight junction proteins claudin 5 and occludin, the efflux transporters P-glycoprotein (PgP) and breast cancer related protein (BCRP), and the transferrin receptor).

No MeSH data available.


Related in: MedlinePlus