Transport Mode
Liquid Nitrogen Transportation
1、Adsorptive Liquid Nitrogen Transportation ,No free liquid nitrogen ,White vapor emission indicates normal operation.
2、Temperature monitoring devices should closely monitor the temperature inside the tank during transportation. If any abnormalities are detected, you can copy the PDF and Excel data from the temperature recorder. (One end of the temperature monitoring device can be unplugged to reveal a USB connector. Simply insert it into a computer to copy the data. If you encounter any difficulties, you can contact the sales staff and technical support for assistance.)
3、Alloy Combination Lock Designed to ensure no third-party access to the LN₂ tank or cell samples during transportation from dispatch to receipt, thereby safeguarding cell integrity.
4、GPS Tracker Enables real-time tracking of cell transportation routes to prevent loss.
5、Liquid Nitrogen Tank, temperature monitoring device, alloy combination lock, and GPS tracker are returnable components. Please store them properly and avoid damage; failure to comply will result in blacklisting.
Reference Citation
blush.
Scope of Application
Human liver sinusoidal endothelial cells (LSECs), as crucial non-parenchymal cells in the liver, have become pivotal research subjects in multiple scientific fields due to their unique structural and functional properties. Below is a summary of their primary research applications:
1. Research on Mechanisms of Liver Diseases
LSECs play central roles in multiple hepatic pathological processes, serving as essential models for investigating hepatic fibrosis, fatty liver disease, hepatocellular carcinoma (HCC), and liver regeneration：
Hepatic fibrosis and liver cirrhosis: Defenestration (capillarization) of liver sinusoidal endothelial cells (LSECs) is an early event in hepatic fibrosis, driving the fibrotic process by activating hepatic stellate cells (HSCs) to secrete collagen.
Fatty Liver Diseases: LSECs modulate the pathological progression of non-alcoholic fatty liver disease (NAFLD) and alcoholic fatty liver (AFL) through lipid metabolism regulation (e.g., STAT3 signaling) and anti-inflammatory functions.
Hepatocellular Carcinoma (HCC): LSECs secrete pro-angiogenic factors (e.g., VEGF) to actively participate in tumor microenvironment remodeling and metastatic regulation, while aberrant expression of their signature proteins (e.g., PLVAP) demonstrates prognostic correlation in HCC clinical outcomes.
Liver Regeneration: LSECs synergize with hepatocytes in secreting growth factors (e.g., Hepatocyte Growth Factor, HGF) to orchestrate post-injury tissue repair and functional restoration through spatially-regulated paracrine signaling.
2. Immune Regulation and Tolerance Mechanisms Research
Liver sinusoidal endothelial cells (LSECs) play dual roles in the hepatic immune microenvironment and serve as key targets for studying immune tolerance and inflammatory balance:
Induction of immune tolerance: By expressing scavenger receptors (such as SR-H) and pattern recognition receptors, LSECs clear pathogens and suppress T cell activation to maintain immune tolerance in the liver, but this may also lead to persistent viral infection or tumor immune escape.
Inflammatory regulation: LSECs secrete anti-inflammatory factors (such as prostaglandins, nitric oxide) to suppress excessive inflammatory responses, while clearing endotoxins and inflammatory mediators from the blood through their endocytic function.
Coordination of the entero-hepatic immune axis: LSECs play a bridging role in gut-derived antigen processing by regulating immune cell migration (e.g., macrophages, T cells), thereby influencing the progression of inflammatory liver diseases.
The highly efficient scavenger function of LSECs renders them valuable in drug delivery and toxicity assessment:
Drug clearance mechanism: LSECs rapidly clear macromolecules (e.g., lipoproteins) and nanoparticles from the blood via scavenger receptors (such as Stabilin-1/2), influencing drug half-life and efficacy—key factors in optimizing nanomedicine design.
Targeted therapy development: Investigating the clearance mechanisms of LSECs can provide strategies to reduce non-specific drug uptake, such as modifying the surface of nanoparticles to evade recognition by LSECs.
Toxicity assessment models: The sensitivity of LSECs to drugs or environmental toxins (such as sinusoidal capillarization) is frequently used to evaluate mechanisms of liver injury and screen for protective drugs.
4. Vascular Biology and Microcirculation Research
The unique vascular properties of LSECs provide a research platform for angiogenesis and microcirculation regulation:
Vascular permeability regulation: The fenestral structure and dynamic changes of LSECs (regulated by the Rho-ROCK pathway) influence substance exchange across liver sinusoids, making them an ideal model for studying mechanisms of vascular permeability.
Coagulation and Anticoagulant Balance: Liver sinusoidal endothelial cells (LSECs) maintain sinusoidal blood flow patency by secreting anticoagulant mediators such as heparin sulfate proteoglycans (HSPGs) and prostacyclin (PGI2). Dysregulation of these functions contributes to prothrombotic complications in liver diseases, including portal vein thrombosis and sinusoidal obstruction syndrome.。
Angiogenesis Research: LSECs drive vascular remodeling processes by secreting pro-angiogenic factors (e.g., Vascular Endothelial Growth Factor [VEGF], Angiopoietin-2), playing dual regulatory roles in both physiological liver regeneration and pathological tumor neovascularization.
5. Biomaterials and Tissue Engineering Applications
The unique biological properties of LSECs hold significant potential for advancing artificial liver support system (ALSS) development and innovating next-generation anticoagulant biomaterials.
Artificial liver devices: The endothelialized surface of LSECs mimics the metabolic and immune barrier functions of natural liver sinusoids, serving as a basis for constructing bioreactors or organ-on-a-chip systems.
Anticoagulant materials: The natural anticoagulant properties of LSECs (such as the secretion of PGI₂) are used in designing antithrombotic coatings to improve the biocompatibility of medical devices.
Conclusion
The research applications of human liver sinusoidal endothelial cells (LSECs) span multiple fields, including mechanisms of liver diseases, immune regulation, drug development, vascular biology, and biomaterials. Their multifunctional properties make them a crucial bridge connecting basic research and clinical translation. Future studies could further explore novel mechanisms of LSECs in metabolic regulation (e.g., vitamin A storage) and interorgan interactions (such as the entero-hepatic axis).
由于我司产品说明书半年更新一次，以下步骤仅作参考，以公司随货说明书为准
II 试剂与材料
Ø 肝窦内皮细胞（Cat# LV- LESC001/2/3/4）
Ø 肝窦内皮细胞培养基（Cat# LV- LSEM001）
Ø 胶原包被培养板/皿/瓶（LV-Coated）
Ø 无菌15ml离心管
Ø 一次性移液管
Ø 37℃/5%CO2培养箱
Ø 移液枪
Ø 恒温水浴锅（37℃预热）
Ø 冷冻水平离心机（带水平转子，可离心15ml离心管）
Ø 生物安全柜
Ø 宽口移液枪头（普通移液枪头剪去尖头，再灭菌使用）
Ø 75%酒精
重要提示：解冻时，冻存细胞转移必须使用液氮转移；在整个复苏实验过程中必须全程使用宽口枪头。
III 细胞的复苏与铺板
1. 肝窦内皮细胞培养基放入37℃恒温水浴锅中充分预热。
2.将冻存的肝窦内皮细胞从冷藏位置经液氮迅速转移至37℃恒温水浴锅中，将冻存管尽可能多的浸入
1. 37℃水中，顺时针水平摇动，但必须确保冻存管管盖保持在水面以上。
2. 解冻冻存管约90~120s，至冻存管中只有少量碎冰漂浮即可。
3. 用75%酒精消毒冻存管，并将其转移到生物安全柜。
4. 用宽口枪头将细胞重悬（轻轻吹打2次），并转移至含10ml预热培养基的15ml离心管中（注意：冻存管与枪头上残留细胞，可用1ml复苏培养基润洗冻存管与枪头）。
5. 将细胞逐滴加入预热培养基中，最后轻微上下颠倒3次混匀。
6. 600g，4℃离心5min，去上清并用培养基重悬。
7. 沉淀的细胞用肝窦内皮细胞培养基重悬并定容至4ml，用台盼蓝排除法测定细胞的活力和细胞总量。
8. 将细胞按3×104cells/cm2接种至胶原包被培养板中。摇匀，在37℃/5%CO2培养箱中培养，24h后换液。
IV 细胞培养与传代
1. 细胞融合度达到80%时可进行传代。
2. 提前将培养基、PBS、胰酶放入37℃水浴锅内预热，用75%酒精擦拭后再放入超净台内。
3. 吸除旧培养液，加少量PBS润洗细胞，加入适量胰酶，使加入的胰酶能盖住细胞，37℃孵育，约2~3min后显微镜下观察。
4. 待贴壁细胞间间隙变大、细胞趋于圆形但还未漂起时弃去胰酶，加入新鲜培养基，晃动细胞板，终止胰酶作用，用吸管小心吹打贴壁的细胞，制成细胞悬液（控制吹打的力度，避免产生大量的气泡）。
5. 600g，室温离心5min，去上清并用肝窦内皮细胞培养基重悬。
6. 将细胞悬液按3×104cells/cm2接种到新的胶原包被培养瓶/板内，置37℃/5%CO2培养箱中培养，隔天观察贴壁生长情况。
