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Melanoma cells homing to the brain: an in vitro model.

Rizzo A, Vasco C, Girgenti V, Fugnanesi V, Calatozzolo C, Canazza A, Salmaggi A, Rivoltini L, Morbin M, Ciusani E - Biomed Res Int (2015)

Bottom Line: M2 cells showed a statistically significant higher capability to pass across the in vitro BBB model, compared to M1.PCR array data showed that M2 had a higher expression of several matrix metalloproteinase proteins (MMPs) compared to M1.Future studies will be necessary to deepen the mechanisms of central nervous system invasion.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Clinical Pathology and Medical Genetics, Foundation IRCCS Neurological Institute Carlo Besta, Via Celoria 11, 20133 Milano, Italy.

ABSTRACT
We developed an in vitro contact through-feet blood brain barrier (BBB) model built using type IV collagen, rat astrocytes, and human umbilical vein endothelial cells (HUVECs) cocultured through Transwell porous polycarbonate membrane. The contact between astrocytes and HUVECs was demonstrated by electron microscopy: astrocytes endfeet pass through the 8.0 μm pores inducing HUVECs to assume a cerebral phenotype. Using this model we evaluated transmigration of melanoma cells from two different patients (M1 and M2) selected among seven melanoma primary cultures. M2 cells showed a statistically significant higher capability to pass across the in vitro BBB model, compared to M1. Expression of adhesion molecules was evaluated by flow cytometry: a statistically significant increased expression of MCAM, αvβ3, and CD49b was detected in M1. PCR array data showed that M2 had a higher expression of several matrix metalloproteinase proteins (MMPs) compared to M1. Specifically, data suggest that MMP2 and MMP9 could be directly involved in BBB permeability and that brain invasion by melanoma cells could be related to the overexpression of many MMPs. Future studies will be necessary to deepen the mechanisms of central nervous system invasion.

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Related in: MedlinePlus

To exclude major differences in cell volume between M1 and M2 cells eventually accounting for the different ability to transmigrate, the forward scatter of M1 (black bar) and M2 (grey bar) cell was measured by flow cytometry. Despite M2 showing a statistically significant increased migration through the in vitro BBB compared to M1, their volume was slightly higher than that of M1 (310 versus 250 arbitrary unit) suggesting that cell volume was not responsible for the differences observed in transmigration.
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fig5: To exclude major differences in cell volume between M1 and M2 cells eventually accounting for the different ability to transmigrate, the forward scatter of M1 (black bar) and M2 (grey bar) cell was measured by flow cytometry. Despite M2 showing a statistically significant increased migration through the in vitro BBB compared to M1, their volume was slightly higher than that of M1 (310 versus 250 arbitrary unit) suggesting that cell volume was not responsible for the differences observed in transmigration.

Mentions: Since the experimental model forces cells to pass through 8 μm pores, we assumed the cell volume as a relevant parameter in our assay. We therefore measured the cellular volume using flow cytometry; forward scatter was measured for both cell lines and results are reported in Figure 5. No statistically significant differences were detected in cellular volume between M1 and M2 cells.


Melanoma cells homing to the brain: an in vitro model.

Rizzo A, Vasco C, Girgenti V, Fugnanesi V, Calatozzolo C, Canazza A, Salmaggi A, Rivoltini L, Morbin M, Ciusani E - Biomed Res Int (2015)

To exclude major differences in cell volume between M1 and M2 cells eventually accounting for the different ability to transmigrate, the forward scatter of M1 (black bar) and M2 (grey bar) cell was measured by flow cytometry. Despite M2 showing a statistically significant increased migration through the in vitro BBB compared to M1, their volume was slightly higher than that of M1 (310 versus 250 arbitrary unit) suggesting that cell volume was not responsible for the differences observed in transmigration.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4321090&req=5

fig5: To exclude major differences in cell volume between M1 and M2 cells eventually accounting for the different ability to transmigrate, the forward scatter of M1 (black bar) and M2 (grey bar) cell was measured by flow cytometry. Despite M2 showing a statistically significant increased migration through the in vitro BBB compared to M1, their volume was slightly higher than that of M1 (310 versus 250 arbitrary unit) suggesting that cell volume was not responsible for the differences observed in transmigration.
Mentions: Since the experimental model forces cells to pass through 8 μm pores, we assumed the cell volume as a relevant parameter in our assay. We therefore measured the cellular volume using flow cytometry; forward scatter was measured for both cell lines and results are reported in Figure 5. No statistically significant differences were detected in cellular volume between M1 and M2 cells.

Bottom Line: M2 cells showed a statistically significant higher capability to pass across the in vitro BBB model, compared to M1.PCR array data showed that M2 had a higher expression of several matrix metalloproteinase proteins (MMPs) compared to M1.Future studies will be necessary to deepen the mechanisms of central nervous system invasion.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Clinical Pathology and Medical Genetics, Foundation IRCCS Neurological Institute Carlo Besta, Via Celoria 11, 20133 Milano, Italy.

ABSTRACT
We developed an in vitro contact through-feet blood brain barrier (BBB) model built using type IV collagen, rat astrocytes, and human umbilical vein endothelial cells (HUVECs) cocultured through Transwell porous polycarbonate membrane. The contact between astrocytes and HUVECs was demonstrated by electron microscopy: astrocytes endfeet pass through the 8.0 μm pores inducing HUVECs to assume a cerebral phenotype. Using this model we evaluated transmigration of melanoma cells from two different patients (M1 and M2) selected among seven melanoma primary cultures. M2 cells showed a statistically significant higher capability to pass across the in vitro BBB model, compared to M1. Expression of adhesion molecules was evaluated by flow cytometry: a statistically significant increased expression of MCAM, αvβ3, and CD49b was detected in M1. PCR array data showed that M2 had a higher expression of several matrix metalloproteinase proteins (MMPs) compared to M1. Specifically, data suggest that MMP2 and MMP9 could be directly involved in BBB permeability and that brain invasion by melanoma cells could be related to the overexpression of many MMPs. Future studies will be necessary to deepen the mechanisms of central nervous system invasion.

Show MeSH
Related in: MedlinePlus