詳細介紹
3D血腦屏障模型芯片,SynBBB,SynBBB 3D Model
SynBBB 3D血腦屏障模型芯片,SynBBB 3D Blood Brain Barrier Model,SynBBB 3D Model – Assay Kits,SynBBB 3D Model Assay Kit SynBBB 3D Model Chip,SynBBB 3D Model Starter Kit
SynBBB 3D Blood Brain Barrier Model – Real-time visualization of cellular and barrier functionality
SynVivo的SynBBB 3D血腦屏障模型通過模擬與跨血腦屏障(BBB)的內皮細胞通訊的腦組織細胞的組織切片來重建體內微環境。剪切誘導的內皮細胞緊密連接在Transwell®模型中無法實現,而在SynBBB模型中使用生理性流體流很容易實現。緊密變化的形成可以使用SynVivo細胞阻抗分析儀通過生化或電氣分析(評估電阻變化)進行測量。腦組織細胞與內皮細胞之間的相互作用在SynBBB分析中很容易觀察到。 Transwell模型不允許實時顯示這些細胞相互作用,這對于了解BBB微環境至關重要。
SynBBB是wei一可以實現以下功能的體外BBB模型:
準確的體內血液動力學切應力
實時可視化細胞和屏障功能
大大減少了成本和時間
穩健易用的協議
BBB模型的示意圖。頂腔(外通道)用于培養血管(內皮細胞),而基底外側腔(中央腔)用于培養腦組織細胞(星形細胞,周細胞,神經元)。多孔結構使血管細胞與組織細胞之間可以進行通訊。
SynBBB系統是一個高度通用的平臺,可用于調查:
緊密連接蛋白:確定緊密連接蛋白的水平,即調節BBB的小帶閉合蛋白,claudins和occludins。
轉運蛋白:分析正常和功能異常的血腦屏障中轉運蛋白的功能(例如Pgp)。
藥物滲透性:評估治療劑和小分子穿過BBB內皮細胞的實時滲透性。
炎癥:了解炎癥反應對血腦屏障調節的潛在機制。
細胞遷移:可視化并量化免疫細胞在BBB中的實時遷移。
滲透性變化:對正常和功能異常的血腦屏障進行基因組,蛋白質組和代謝分析。
神經毒性:分析化學,生物和物理試劑對血腦屏障細胞的毒性作用。
神經腫瘤學:研究腫瘤細胞對血腦屏障的影響。
根據您的研究需求,您可以從“基本” SynBBB模型或“ TEER兼容” SynBBB配置中進行選擇。
SynBBB 3D模型套件組件
可以以試劑盒形式購買運行SynBBB分析所需的所有基本組件。 根據個人研究需求,您可以從SynBBB芯片的“基本”或“ TEER兼容”配置中進行選擇。 包括所有附件,包括管子,夾子,針頭和注射器。 入門工具包還將包括氣動啟動裝置(運行SynBBB分析所需)和細胞阻抗分析儀(收集SynBBB TEER測量值所需)。
套件內容和說明
SynBBB Kits and Chips | Basic BBB Assay Kit Cat# 402001 | Basic BBB Starter Kit Cat# 402002 | BBB-TEER Assay Kit Cat# 402003 | BBB-TEER Starter Kit Cat# 402004 |
102005-SB Chips (3uM slit) (10) | ? | ? | ||
102015-SB Chips (3uM slit-Teer compatible) (10) | ? | ? | ||
Pneumatic Primer and Adapter | ? | ? | ||
Manifold (5 port) | ? | ? | ||
Blunt Tip Needles 0.5” long, 24ga (50) | ? | ? | ? | ? |
Tygon Tubing 0.2” ID x 0.6” OD (100 ft) | ? | ? | ? | ? |
1 mL Syringes (50) | ? | ? | ? | ? |
Slide Clamps (25) | ? | ? | ? | ? |
Impedance Analyzer | ? | |||
Electrodes (20) | ? | ? |
SynVivo used to create the first neonatal BBB model on a chip
Researchers at Temple University used the SynVivo® SynBBBTM cell-based in vitro assay platform to model the attributes and functions of the neonatal stage blood-brain barrier (BBB) [1]. The SynBBB model closely mimics the in vivo microenvironment including three-dimensional morphology, cellular interactions and flow characteristics on a microfluidic chip. This work marks the first dynamic in vitro neonatal BBB model that offers real time visualization and analysis and is suitable for studies of BBB function as well as screening of novel therapeutics.
“The work is important because studies of neonatal neuropathologies and development of appropriate therapeutics are hampered by a lack of relevant in vitro models of the neonatal blood-brain barrier,” said Dr. Sudhir Deosarkar, the lead author of this paper.
In the SynBBB assay, which includes a tissue compartment and vascular channels placed side-by-side and separated by an engineered porous barrier, the researchers were able to co-culture neonatal rat brain endothelial cells and rat astrocytes under physiological conditions observed in vivo. The endothelial cells formed a full lumen and exhibited tight junction formation which increased under co-culture with astrocytes. The permeability of small molecules in the developed model was found to in excellent agreement with in vivo observations.
“The real-time visualization capabilities of the SynBBB co-culture platform allowed, for the first time, visualization of astrocyte end-feet and endothelial cell interactions in anin vitro model,” said Prof. Mohammad Kiani who is the senior author of the paper. “This is a unique capability and will help us to understand and develop therapeutics for several developmental disorders and diseases of the brain.”
The PLOS ONE paper shows that in contrast to transwell models, the SynBBB model exhibits significantly improved barrier characteristics similar to in vivo observations.
1A Novel Dynamic Neonatal Blood-Brain Barrier on a Chip. S. Deosarkar, B. Prabhakarpandian, B. Wang, J.B. Sheffield, B. Krynska, M. Kiani. PLOS ONE, 2015, DOI: 10.1371/journal.pone.0142725
The SynBBB 3D model has been validated in various BBB Assays
Mono-Culture Assays
Shear-induced endothelial cell tight junctions, which cannot be achieved in the Transwell® model, are easily achieved in the SynBBB assay using fluid perfusion. Formation of tight changes can be measured using biochemical or electrical analysis (assessing changes in electrical resistance) with the SynVivo Cell Impedance Analyzer.
Primary endothelial cells are cultured in the vascular channel under physiological fluid flow. Cells are stained for tight junction markers highlighting the increase under fluid flow compared to static conditions. The Cell Impedance Analyzer system is used to measure increases in Ohmic resistance (TEER), associated with the formation of tight junctions.
Top Left Panel: Phase Contrast imaging of brain endothelial cells cultured in the SynBBB model. Bottom Left Panel: Calcein AM and Ethidium homodimer-1 labeled brain endothelial cells indicating a highly viable population of cells in the SynBBB model. Right Panel: Plot highlighting the importance of flow on brain endothelial cells with increased TEER.
Co-Culture with Tissue Cells
Interactions between brain tissue cells and endothelial cells are readily visualized in the SynBBB assay. Transwell models do not allow real-time visualization of these cellular interactions, which are critical for understanding of the physiological environment.
Endothelial cells are cultured under flow in the vascular channel, and the tissue chamber is cultured with primary brain cells, such as astrocytes. Increases in Ohmic resistance across the barrier, measured with the Cell Impedance Analyzer, are associated with tight junction formation across the BBB. Endothelial cells co-cultured with astrocytes form significantly tighter cell junctions compared to mono-cultured endothelial cells.
Left Panel: CD-31 (green) stained endothelial cells and GFAP (red) stained astrocytes. All nucleus are stained with DAPI (blue). Right Panel: Plot highlighting increased TEER with co-culture of endothelial cells and astrocytes.
Real-Time Permeability Assays
Unlike BBB models which are arranged in top to bottom architecture (i.e., Transwell), small molecule transport can be assessed and quantified in real-time across the SynBBB system due to its side-by-side architecture.
A fluorescently-labeled drug molecule of interest is perfused through the vascular channels at physiological flow rate. Real-time videos are acquired and analyzed to calculate the rate of permeability into the tissue chamber. Different rates of permeability is observed across the BBB due to tight junctions of endothelial cells.
Time-lapse imaging of permeability of small molecules across a tightly formed BBB.
Time-lapse imaging of permeability of small molecules across a leaky BBB.
Real-Time Tight Junction Modulation
SynBBB can be used to model inflammation responses. A pro-inflammatory compound, such as TNF-α, is added to mono-cultured endothelial cells to modulate the tight junctions, followed by a period of recovery under perfusion flow. Electrical resistance measurements provide a non-invasive method for real-time monitoring of tight junctions.
Modulation of Inflammation responses in SynBBB model. TNF-alpha induced leakiness in the BBB measured by changes in the resistance across the endothelial cells. Removal of TNF-alpha followed by media perfusion under physiological flow conditions enables recovery of the tight junction leading to increased tight junction formation. Static cells maintain a constant resistance due to lack of tight junctions.
SynBBB 3D模型–入門套件
包括消耗品(10個芯片,百英尺管,25個滑動夾具,50個鈍頭針頭和50個1毫升注射器)。 入門套件還將包括氣動啟動裝置(接種細胞所需)和細胞阻抗分析儀(僅適用于TEER配置)。
貨號:402002,SynBBB 3D模型入門套件(基本
貨號:402004,SynBBB 3D模型入門套件(TEER
SynBBB 3D模型–檢測套件
包括消耗品(10個芯片,百英尺管,25個滑動夾具,50個鈍頭針頭和50個1毫升注射器)。
貨號:402001 SynBBB 3D模型測定試劑盒(基本配置)
貨號:402003 SynBBB 3D模型測定套件(TEER配置)
SynBBB 3D模型–芯片
貨號:102005-SB SynBBB 3D模型芯片(基本配置)
貨號:102015-SB SynBBB 3D模型芯片(TEER配置)