R&D Products

 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

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Scope of Application

Hepatic stellate cells (HSCs) hold significant value in biomedical research, and their applications can be summarized into the following core directions:

1.Mechanisms of Liver Fibrosis and Cirrhosis

Hepatic stellate cells (HSCs) are the central effector cells in liver fibrosis. Under pathological conditions, HSCs are activated and transdifferentiate into myofibroblasts, secreting excessive extracellular matrix (ECM) components (e.g., collagen I/III) that drive fibrogenesis. Researchers investigate the molecular mechanisms of fibrosis by: In vitro models: Utilizing activated HSCs (e.g., the human LX-2 cell line) to analyze cytokine secretion (e.g., TGF-β, IL-6), ECM composition, and signaling pathways (e.g., AKT phosphorylation).

Genetic models: Employing gene-knockout animals (e.g., Plvap-null mice) to dissect HSC-specific molecular networks.

Functional studies: Characterizing HSC contractility to elucidate mechanisms of sinusoidal hypertension in cirrhosis.

2. Research on Metabolic Regulation and Energy Balance

Hepatic stellate cells (HSCs) regulate hepatic metabolic substrate preferences through specific proteins such as PLVAP (Plasmalemma Vesicle-Associated Protein). For instance, during fasting, PLVAP in HSCs modulates fatty acid esterification and oxidation, determining whether the liver prioritizes fatty acids or carbohydrates for energy production. PLVAP-knockout mice exhibit enhanced glucose metabolism capacity and activated insulin signaling pathways, offering novel insights into metabolic disorders such as diabetes and non-alcoholic fatty liver disease (NAFLD). Furthermore, the vitamin A storage capacity of HSCs has been leveraged to study lipid-soluble vitamin metabolism and its dysregulation in chronic liver diseases.

3. Models of Liver Regeneration and Cell-Cell Interactions

Hepatic stellate cells (HSCs) secrete growth factors (e.g., HGF and VEGF) to promote hepatocyte proliferation and angiogenesis, playing a dual role in liver injury repair. In research, co-culture systems (e.g., HSCs with hepatocytes or liver progenitor cells) are widely used to mimic the regenerative microenvironment. These models enable analysis of HSC-mediated effects on hepatocyte functions, such as albumin secretion and drug-metabolizing enzyme activity (e.g., CYP450 enzymes). Additionally, such systems help elucidate interaction mechanisms between HSCs and immune cells (e.g., macrophages), dissecting the balance between inflammation and regeneration during liver repair.

4. Drug Development and Anti-fibrotic Therapy Screening

Immortalized HSC cell lines (e.g., LX-2, HSC-Li) are indispensable tools for anti-fibrotic drug screening. Researchers evaluate candidate compounds through in vitro assays that measure their inhibitory effects on HSC proliferation, α-smooth muscle actin (α-SMA) expression, and collagen secretion, followed by in vivo validation in animal models (e.g., CCl₄-induced fibrosis). For instance, small-molecule inhibitors targeting HSC activation pathways (e.g., TGF-β/Smad signaling, PDGF receptor tyrosine kinase) and gene therapies (e.g., siRNA against Col1a1) have advanced to the preclinical research stage.

5. Research on Tumor Microenvironment and Cancer Progression

Activated hepatic stellate cells (HSCs) participate in the remodeling of the liver cancer microenvironment and the invasion/metastasis of tumor cells by secreting matrix metalloproteinases (MMPs) and inflammatory factors. In scientific research, co-culture models of HSCs and liver cancer cells are frequently utilized to investigate their regulatory roles in tumor growth, angiogenesis, and immune escape, thereby providing new strategies for liver cancer treatment.

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 肝星形细胞（Cat# LV-HSC001/2/3/4）              

Ø 复苏培养基（Cat#LV-Rec001）

Ø 肝星形细胞培养基（Cat# LV-HSCM001）

Ø 胶原包被板/皿/瓶（Cat# 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. 复苏培养基放入37℃水浴锅中充分预热，铺板培养基放入37℃恒温水浴锅中充分预热。

2. 将冻存的肝星形细胞从冷藏位置经液氮迅速转移至37℃恒温水浴锅中，尽可能多的浸入37℃水中，

1. 顺时针水平摇动，但必须确保冻存管盖子保持在水面以上。

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复苏培养基清洗冻存管与枪头）。

5. 将细胞悬液逐滴加入预热复苏培养基，每1ml细胞悬液加入10ml复苏培养基（注意：前3ml要缓慢逐滴加入并轻微摇晃，后面7ml可加快速度），最后轻微上下颠倒1次混匀。

6. 600g，4℃离心5min，去上清并用培养基重悬。

7. 沉淀的细胞用肝星形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，置37℃/ 5% CO₂培养箱中培养，24小时后换液。

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%酒精擦拭后再放入生物安全柜内。

3. 吸除旧培养液，加少量PBS润洗细胞，弃PBS后加适量胰酶，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。

Cells should not be re-cryopreserved after thawing.

VI Regarding After-Sales Service

      Quality Assurance & After-Sales Policy In the event of product quality issues, please: Collect original data Contact our sales representatives or technical support team immediately We will dispatch personnel to address the issue promptly.

      After-Sales Validity Period & Submitted Original Data：

      Resuscitation issues: Report any abnormalities immediately and provide results of Trypan blue staining or AO/PI staining.

      Contamination issues: Report within 96 hours after resuscitation and provide phase-contrast microscope photos.

      Purity issues: Report within one month of detection and provide immunofluorescence or flow cytometry results.

VII  Contact Number

      Company Telephone：0755-28284050

      Technical Support：19902901483（ Dr. Zhou）