體外血腦屏障模型裝置-體外3D細(xì)胞屏障培養(yǎng)

flocel體外血腦屏障實(shí)驗(yàn)?zāi)Ｐ拖到y(tǒng)

—Flocel體外三維動態(tài)血腦屏障（Dynamic In Vitro Blood-Brain Barrier Model）

Blood-Brain Barrier Model）

體外血腦屏障測量模型一般包括：

1）跨細(xì)胞膜電阻測量（TEER 檢測）系統(tǒng)

2）計(jì)算機(jī)控制的脈動泵

3）flocel控制軟件

是Flocel公司生產(chǎn)和銷售的一個更實(shí)際的、動態(tài)的體外血腦屏障（DIV-BBB™）模型，更準(zhǔn)確地反映真正的血腦屏障特性。

迄今為止，研究者已經(jīng)使用靜態(tài)BBB模型或動物模型進(jìn)行了測試。和這些方法對比，flocel的DIV-BBB模型的優(yōu)勢：

更的藥代動力學(xué)和毒理學(xué)研究

● 更準(zhǔn)確地反映體內(nèi)血腦屏障特性

● 模仿重要的內(nèi)皮細(xì)胞-星形膠質(zhì)細(xì)胞相互作用

● 電測量血腦屏障的完整性

● 能使用真實(shí)的人體細(xì)胞

● 形成比現(xiàn)有靜態(tài)模型更緊密的連接

● 可大幅降低藥物開發(fā)成本

特點(diǎn)

與傳統(tǒng)血腦屏障系統(tǒng)相比，F(xiàn)locel體外動態(tài)血腦屏障系統(tǒng)模型特點(diǎn)如下：

• 可以做更準(zhǔn)確的藥代動力學(xué)和毒物學(xué)的研究

• 更清楚準(zhǔn)確的反應(yīng)了體內(nèi)血腦屏障的特性

• 模擬重要的內(nèi)皮細(xì)胞與星形膠質(zhì)細(xì)胞之間的作用

• 用電力方法檢測了血腦屏障的完整性

• 可以用真實(shí)的細(xì)胞做實(shí)驗(yàn)

• 比現(xiàn)存的靜態(tài)模型更能形成緊密的連接

• 降低藥物研發(fā)的成本

1.DIV-BBB™ 管腔

動態(tài)血腦屏障參數(shù)

2.跨細(xì)胞膜電阻測量（TEER 檢測）系統(tǒng)

TEER檢測系統(tǒng)為研發(fā)人員提供了一個快速、簡便地評估血腦屏障完整性方法，此外，電阻抗值（>1000 Ω-cm）也非常接近體內(nèi)數(shù)值。TEER檢測系統(tǒng)同時配帶了所需檢測軟件、電線和襯板，可供4個DIV-BBB管腔同時使用。其中檢測軟件可以自定義設(shè)置泵的流速、頻率、波形等參數(shù)。其它特性及優(yōu)點(diǎn)見下表：

- Transcending the Transwell -

______________________________________

"...the exposure to intraluminal flow plays an essential role in promoting endothelial cell differentiation and increasing BBB tightness..."

"...in contrast to the Transwell apparatus...endothelial cells grown under dynamic conditions ...develop in a monolayer fashion, thus mimicking the structural physiology of capillaries in vivo..."

Side by side comparison between dynamic versus static models
of blood–brain barrier in vitro: A permeability study

Janigro et al., 2006

______________________________________

Our mission at Flocel Inc. is not only to to provide high-quality products and services for in vitro 3D cell barrier culture, but also to convey -with integrity and transparency- the advantages of our DIV-BBB 2.0 system compared to other approaches --such as the Transwell.

- Stronger Barriers, Stronger Data -

A comparative study between cell barriers cultured in a static flow (i.e., Transwell) and dynamic flow (i.e., DIV-BBB) environment showed that fluid flow conditions greatly influence how a cell barrier develops. Trans-endothelial electrical resistance (TEER) across the DIV-BBB culture (B) increased to values 10-fold higher than those recorded in the Transwell (A), and much closer to values observed in vivo. This is because dynamic flow causes shear stress on the apical surface of the cell, and influences cell development and function.

This concept is reiterated in the inset microscope images of the figure above: A cell culture that has been subjected to dynamic flow and physiologically-representative levels of shear stress (B) resulted in a confluent monolayer of cells expressing tight junctions. The cell barrier in under static flow (A) receives no shear stress, and instead forms a multilayer of cells with less tight junction organization.

- Higher Selective Permeability -

Owing to the stronger and more physiologically-accurate cell barrier grown under dynamic flow conditions, the DIV-BBB also presents a more stringent regulation of selective permeability. Tests conducted between cells cultured in the DIV-BBB (C) and Transwell (D) for passage of phenytoin (an epileptic drug with in vivo permeability levels of 0.0001 cm/s) showed markedly higher levels of permeability through the Transwell than the DIV-BBB, and 4-fold higher than in vivo. The stronger and more physiologically-accurate cell barriers grown under the dynamic fluid conditions of the DIV-BBB allow for more accurate testing of drug permeability and pharmacokinetics.

- What if I already own Transwell products? -

If you would prefer to continue using your Transwell products (such as Endohm-6, Endohm-12, Endohm-24SNAP cell culture chambers or any "chopstick" electrodes), we also offer a Trans-endothelial Electrical Resistance (TEER) Measurement Unit that is compatible with these Endohm products, and much more versatile! Not only does our TEER Measurement Unit allow for frequency sweeps between 0.1 Hz and 1,000 Hz (0.1 Hz sampling resolution), it can be directed by the user to automatically sample single-frequency and frequency sweep data for an indefinite amount of time, and save this data to a .xls or .csv save file. No  more pushing a button and jotting down the TEER value every time you want to take a measurement!

測試服務(wù)

世聯(lián)博研和Flocel Inc.聯(lián)合，提供以合同為基礎(chǔ)的體外 BBB滲透性測試。費(fèi)用結(jié)構(gòu)由待測化合物的數(shù)量和測試的復(fù)雜性決定。這項(xiàng)服務(wù)在克利夫蘭基金會，斯坦福大學(xué)和歐洲的實(shí)驗(yàn)室進(jìn)行。我們還提供腦生物標(biāo)志物的咨詢服務(wù)和測試儀器，并通過分析外周血檢測人體受試者血腦屏障的完整性，提供診斷患者或運(yùn)動員血腦屏障的方法。
輕型顱腦損傷（MHI和MTBI）占顱腦損傷的95%；診斷和早期管理包括計(jì)算機(jī)斷層掃描（CT）或住院觀察，但是這些選項(xiàng)都不理想。在醫(yī)院觀察是不經(jīng)濟(jì)的；CT有時是不切實(shí)際的，涉及潛在的有害電離輻射。GFAP和S100等腦蛋白在體液中表達(dá)水平與許多神經(jīng)系統(tǒng)疾病有關(guān)聯(lián)性。Flocel開發(fā)了商業(yè)化的血液測試小型工具，給創(chuàng)傷性腦損傷進(jìn)行前期及時診斷，防止嚴(yán)重的延遲性后果。

發(fā)表文獻(xiàn)：

- Publications -

• The role of shear stress in blood-brain barrier endothelial physiology.
       Cucullo L, Hossain M, Puvenna V, Marchi N, Janigro D.

• A dynamic in vitro BBB model for the study of immune cell trafficking into the central nervous system.
       Cucullo L, Marchi N, Hossain M, Janigro D.

• Pattern of P450 expression at the human blood-brain barrier: roles of epileptic condition and laminar flow.
       Ghosh C, Gonzalez-Martinez J, Hossain M, Cucullo L, Fazio V, Janigro D, Marchi N.

• Tobacco smoke: a critical etiological factor for vascular impairment at the blood-brain barrier.
       Hossain M, Sathe T, Fazio V, Mazzone P, Weksler B, Janigro D, Rapp E, Cucullo L.

• Immortalized human brain endothelial cells and flow-based vascular modeling: a marriage of convenience for rational neurovascular studies.
       Cucullo L, Couraud PO, Weksler B, Romero IA, Hossain M, Rapp E, Janigro D.

• Side by side comparison between dynamic versus static models of blood-brain barrier in vitro: a permeability study.
       Santaguida S, Janigro D, Hossain M, Oby E, Rapp E, Cucullo L.

• Glycerophosphoinositol and dexamethasone improve transendothelial electrical resistance in an in vitro study of the blood-brain barrier.
       Cucullo L, Hallene K, Dini G, Dal Toso R, Janigro D.

• A new dynamic in vitro model for the multidimensional study of astrocyte-endothelial cell interactions at the blood-brain barrier.
       Cucullo L, McAllister MS, Kight K, Krizanac-Bengez L, Marroni M, Mayberg MR, Stanness KA, Janigro D.

- Posters -

• A Dynamic Humanized drug resistant In Vitro Blood-Brain Barrier Model to assess the permeability of relevant CNS drugs

• Development of a novel blood-brain barrier model by coupling Immortalized human brain endothelial cells with flow-based cell culture technologies

• Glioblastoma Cell Line Metabolism Under Dynamic and Static Conditions: Effects on tumor growth and BBB integrity

• Humanized in vitro blood-brain barrier models to screen for brain penetration of antiepileptic drugs

• Effect of Shear stress on BBB endothelial cells: A proteomic study by 2-dimensional protein electrophoresis