Mouse Hepatic Stellate Cells (Primary cell immortalization)
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Product Introduction
| Cell Name | Mouse Hepatic Stellate Cells (Primary cell immortalization) |
| Species | Mouse |
| Tissue | Liver tissue |
| Growth Properties | Adherent |
| Morphology | Spindle-shaped; polygonal |
| Isolation Method | Mouse hepatic stellate cells (primary cell immortalization) from Procell Laboratory were generated by lentiviral delivery of the SV40T gene into primary mouse hepatic stellate cells, followed by antibiotic selection and serial subculture screening. Each vial contains approximately 5 × 10⁵ cells. |
| Identity Markers | Mouse hepatic stellate cells (primary cell immortalization) isolated from Procell Laboratory exhibit positive immunofluorescence staining for α-SMA, with a purity greater than 90%. In addition, the cells test negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. |
| Background | Mouse hepatic stellate cells (primary cell immortalization) from Procell Laboratory were isolated by lentiviral transduction of primary mouse hepatic stellate cells with the SV40T gene, followed by antibiotic selection and subculture screening. Mouse hepatic stellate cells are isolated from liver tissue. The liver is an organ primarily responsible for metabolic functions, including detoxification, glycogen storage, and synthesis of secretory proteins. It also produces bile for the digestive system. The liver is the largest internal organ of the body, located in the upper right quadrant of the abdomen, beneath the diaphragm, anterior to the right kidney, and above the stomach. It is the largest digestive gland in the body, appearing as a reddish-brown V-shaped organ. The liver is a major site of urea synthesis and plays a critical role in metabolism. Its position and morphology: the liver lies in the right upper abdomen, hidden under the diaphragm and rib cage. Most of the liver is covered by the costal margin, with only a small area exposed in the epigastric region and right costal margin, directly contacting the anterior abdominal wall. The superior surface of the liver contacts the diaphragm and the anterior abdominal wall. Hepatic stellate cells (HSCs; also known as fat-storing cells, vitamin A-storing cells, perisinusoidal cells, or Ito cells) are the primary source of extracellular matrix (ECM) in the liver. Upon activation, HSCs can further transdifferentiate into myofibroblast-like cells. Various factors that lead to liver fibrosis ultimately target HSCs. Under normal conditions, HSCs are quiescent. When the liver is injured, such as by inflammation or mechanical stress, HSCs become activated. Activated HSCs contribute to liver fibrosis and the remodeling of hepatic structure through proliferation and secretion of ECM components – a central event in liver fibrogenesis. HSCs reside in the space of Disse and are the main source of ECM synthesis and secretion, playing a pivotal role in the development of liver fibrosis. Hepatic stellate cells are liver-specific mesenchymal cells, accounting for approximately 8–13% of all liver cells. As the major ECM-producing cell population in the liver, HSCs not only secrete ECM components such as proteoglycans and glycoproteins, and synthesize a certain amount of collagenase to maintain the normal basement membrane structure, but also regulate sinusoidal microcirculation through the contraction of their processes. Furthermore, HSCs promote hepatocyte proliferation and liver regeneration by synthesizing hepatocyte growth factor (HGF), insulin-like growth factor (IGF), epidermal growth factor (EGF), and other factors. |
| Instructions | 1. Check all containers for leakage or breakage. 2. Remove the frozen cells from the dry ice packaging and immediately transfer them to liquid nitrogen (liquid or vapor phase) for long-term cryopreservation. 3. For the first subculture, a 1:2 split ratio is recommended. Subculture at a 1:2 split ratio; cells can be subcultured for more than 10 passages. Accutase™ digestion is recommended. If trypsin digestion is used, partial cell damage and death may occur. |
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| Reagent Preparation |
The culture period of Mouse Hepatic Stellate Cells (Primary cell immortalization) in vitro is limited; it is recommended to use the dedicated growth medium provided by Procell and follow the correct operation methods for cultivation to ensure the optimal culture state of these cells.
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| Incubation Atmosphere | Air，95%；CO₂，5% | ||||||||
| Temperature | 37℃ | ||||||||
| Subcultivation Ratio | First passage at 1:2 ratio, subsequent passages at 1:2 to 1:6 ratio | ||||||||
| Medium Renewal | Every 2 to 3 days | ||||||||
| Population Doublings | Guaranteed to further expand for 250 population doublings | ||||||||
| Dissociation Reagent | Accutase | ||||||||
| Freeze Medium | General Freezing Medium [PB180436] | ||||||||
| Storage Conditions | For long-term cryopreservation, cryovials should be stored in liquid nitrogen at −150°C to −196°C. Storage at −80°C is restricted to short-term interim use only. |
Documents
Publications
Journal: JOURNAL OF MEDICINAL CHEMISTRY (2026) IF: 6.8
DOI: 10.1021/acs.jmedchem.5c03055
Product Cited: LX-2 Cells Line, Mouse Hepatic Stellate Cells (Primary cell immortalization)
Journal: PHYTOMEDICINE (2025) IF: 8.3
DOI: 10.1016/j.phymed.2025.157572
Product Cited: Mouse Hepatic Stellate Cells (Primary cell immortalization)
FAQs
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Q:1 The color of cryopreserved cells varies, with some appearing pink and others yellowish. Does this variation affect their viability after thawing?
AnswerThis phenomenon frequently occurs between different cryopreservation batches, and is associated with cell density, cryopreservation solution composition, and pH changes induced by temperature variations. This discoloration occurs only occasionally, and does not necessarily indicate compromised cell viability. We recommend attempting cell resuscitation first to assess their condition.
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Q:2 How can I improve the survival rate of cryopreserved cells stored at -80°C?
AnswerWhen storing cryovials, place them in the innermost compartment of the refrigerator's freezer section; avoid placing them on the door shelves or near the refrigerator opening. Additionally, we recommend using refrigerators that are opened less frequently to minimize temperature fluctuations caused by opening and closing the door.
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Q:3 How long can cells be preserved in a -80°C freezer?
AnswerThe preservation duration of cells at -80°C depends on the operating frequency of the refrigerator and the temperature sensitivity of the cells themselves; therefore, there is no standardized storage period. It is recommended to transfer the cells into liquid nitrogen for long-term storage as early as possible.
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Q:4 For frozen cells transported via dry ice, should they be stored at -80°C or in liquid nitrogen upon receipt?
AnswerIt is recommended to immediately transfer the received frozen cells into liquid nitrogen for long-term storage; storage at -80℃ should only be used as a short-term interim solution. This is because at -80℃, cellular metabolism is significantly slowed but not completely halted. In addition, frequent opening and closing of the -80℃ freezer causes temperature fluctuations that reduce cell viability.
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Q:5 My cells are growing very slowly. What could be the cause of this?
AnswerPlease see some common reasons below, and solutions to the issue: - Growth medium is not correct: Use pre-warmed growth medium as recommended by the supplier. - Serum in the growth medium is of poor quality: Use serum from a different lot. and choose good quality serum to ensure nutrition. - Cells have been passaged too many times: Use healthy, low-passage number cells. - Cells were allowed to grow beyond confluency: Passage mammalian cells when they are in the log-phase before they reach confluency. - Culture is contaminated with mycoplasma: Discard cells, media, and reagents. Obtain new stocks of cells, and use them with fresh media and reagents.
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Q:6 What factors can contribute to rapid cell death/culture failure?
AnswerThere are a number of events that can contribute to this: 1. Incorrect CO2 levels:Monitor the level of CO2 manually with a Fyrite kit, available from Bacharach. Check if the manual readings concur with the readings displayed on the incubator. If the incubator has a trace readout, check the printout for fluctuations in CO2 level. Check the settings to ensure that CO2 levels are set at appropriate levels for your cell line (usually between 5 and 10%). Check line connections frequently for leaks. Avoid frequent opening and closing of incubator doors. 2. Temperature fluctuations in the incubator:Monitor the temperature of incubator with a good thermometer inside the incubator. 3. Amphotericin B or other preventive antibiotics/antimycotics are present at toxic concentrations:Use at recommended levels. 4. Humidity is incorrect:Check the water level in the water pan. Humidity is vital to appropriate gas exchange for many types of cells and media. 5. Incorrect osmotic pressure in medium:Check osmolality of complete medium. Most mammalian cells can tolerate an osmolality of 260 to 350 mOsm/kg. Additions of reagents such as HEPES and drugs may affect osmolality. 6. Contamination by microorganisms:Bacterial and fungal contaminations are usually easily visible; symptoms of mycoplasma contamination are more subtle, and careful monitoring of culture morphology and regular testing are necessary to detect this type of contamination. 7. Inappropriate medium is being used:Double-check that the medium used is appropriate for your cell type and culture application. For example, ensure that the medium being used for serum-free culture is actually designed for serum-free culture; make sure that appropriate selective drugs are used at appropriate levels; check the expiration dates for the reagents being used; and store medium at appropriate temperatures in the dark.
