Mouse Ovarian Granulosa Cells (Primary cell immortalization)
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CP-M050Y
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Product Introduction
| Cell Name | Mouse Ovarian Granulosa Cells (Primary cell immortalization) |
| Species | Mouse |
| Tissue | Ovarian tissue |
| Growth Properties | Adherent |
| Morphology | Fibroblast-like |
| Isolation Method | Mouse ovarian granulosa cells (primary cell immortalization) from Procell Laboratory were generated by lentiviral delivery of the SV40T gene into primary mouse ovarian granulosa cells, followed by antibiotic selection and serial subculture screening. Each vial contains approximately 5 × 10⁵ cells. |
| Identity Markers | Mouse ovarian granulosa cells (primary cell immortalization) isolated from Procell Laboratory exhibit positive immunofluorescence staining for FSHR, with a purity greater than 90%. In addition, the cells test negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. |
| Background | Mouse ovarian granulosa cells (primary cell immortalized) are generated from primary mouse ovarian granulosa cells. Primary cells are transduced with the SV40T gene through lentiviral infection, followed by antibiotic screening and continuous passaging to establish a stably immortalized cell line. Primary mouse ovarian granulosa cells are isolated from ovarian tissue. The ovary is the core female reproductive organ responsible for oocyte generation and steroid hormone synthesis. In mice, ovarian functional laterality is distinct, with the left ovary generally exhibiting more stable and superior physiological function, while the right ovary may present partial regression. The ovarian surface presents a grape-like appearance covered with numerous follicles at diverse developmental stages, showing an overall yellowish color and abundant superficial vascular distribution. Ovarian morphological size dynamically changes with age and estrous cycle progression. Notably, ovarian laterality varies across species: most vertebrates have paired functional ovaries, some fish exhibit fused bilateral ovaries, and all birds retain only a functional left ovary. In mammals, ovaries are paired oval organs symmetrically distributed on both sides of the uterus. The ovarian surface is covered by a layer of germinal epithelium, with a thin underlying connective tissue layer. Histologically, the ovarian parenchyma is structurally divided into two distinct regions: the outer cortex and the inner medulla. The ovarian cortex constitutes the main functional region, predominantly composed of developing ovarian follicles and supportive connective tissue. The central medulla consists of loose connective tissue enriched with abundant blood vessels, lymphatic vessels and nerve fibers, providing nutritional and neural support for ovarian tissue. Ovarian granulosa cells are the predominant functional cell population within ovarian follicles. Their proliferation, differentiation and functional status directly govern key ovarian physiological processes, including follicle activation, follicular development, ovulation, corpus luteum formation and steroid hormone secretion. Rapid proliferation and growth of granulosa cells serve as a critical hallmark of normal follicular development. In adult animals, granulosa cell apoptosis is the primary cause of follicular atresia, particularly during the late stages of follicle maturation. In addition, granulosa cells coordinate with theca cells to synthesize and secrete ovarian steroid hormones, and construct a stable microenvironment to support oocyte growth and maturation. Morphologically, ovarian granulosa cells are typically round or oval under microscopic observation. (Note: The description of mouse ovarian laterality indicates predominant left ovarian functionality in rodents, rather than exclusive unilateral development; both ovaries retain physiological functions and participate in reproductive activities.) |
| 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. These cells proliferate rapidly; the split ratio can be adjusted based on actual proliferation status. |
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| Reagent Preparation |
The culture period of Mouse Ovarian Granulosa 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 | 0.25% trypsin | ||||||||
| 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
Feedback FormFAQs
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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.
