TM3 Cell Line

Brief Introduction
Product List

General Information
Organism	Mus musculus, mouse
Cell Line Description	TM3 is a spontaneously immortalized mouse testicular cell line established by Jennie P. Mather in 1980. The cell line was derived from primary cultures enriched in Leydig cells, obtained from the normal testes of 11- to 13-day-old male BALB/c mice (specifically of the BALB/c nu/+ genotype). TM3 cells exhibit an epithelial-like morphology, are anchorage-dependent, and grow as a tightly adherent monolayer. In the fields of reproductive biology and endocrinology, TM3 is recognized as the "gold standard" non-tumorigenic in vitro model for studying Leydig cells. The cell line retains key functional characteristics of Leydig cells, such as specific hormone-stimulated metabolic and signal transduction pathways. It is widely used as a standard model to investigate the microenvironment of spermatogenesis, Leydig cell development, steroid hormone synthesis pathways, and the effects of environmental endocrine disruptors.
Tissue	Testis; Leydig cells
Cell Type	Leydig cell (Testicular interstitial cell)
Strain	BALB/c
Disease	Normal / Non-malignant tissue origin (Spontaneously immortalized)
Morphology	Epithelial-like
Gender	Male
Age	11 to 13 days
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 (Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.)
Applications	1. A highly cited in vitro model for studying the endocrine, paracrine, and autocrine regulatory mechanisms governing Leydig cell function and hormone synthesis. 2. A primary target model in reproductive toxicology for screening the effects of exposure to heavy metals, plasticizers, and environmental endocrine-disrupting chemicals. 3. Used to investigate androgen biosynthetic pathways and cholesterol metabolism under luteinizing hormone (LH) stimulation. 4. Used to elucidate signal transduction pathways involving epidermal growth factor (EGF), luteinizing hormone (LH), and steroid hormone receptors in testicular biology. 5. A host system for evaluating cellular transport mechanisms, lipid-related signaling cascades, and intercellular communication.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	No, non-tumorigenic
Transformant	None; spontaneously immortalized during serial cultivation and selective expansion of Leydig cell-enriched populations.
Hormone Responsiveness	Responds specifically to luteinizing hormone (LH) or human chorionic gonadotropin (hCG) with a robust increase in cyclic AMP (cAMP) production. Conversely, TM3 exhibits no response to follicle-stimulating hormone (FSH), which serves as a physiological marker distinguishing it from Sertoli cells. (Note: The maintenance of high-sensitivity responsiveness to LH is dependent on serum lot selections).
Secretory & Synthesis Profile	1. Cholesterol metabolism: In the presence of luteinizing hormone (LH), these cells are fully capable of actively metabolizing cholesterol. 2. Secretion: Capable of synthesizing and secreting prostaglandin F2α.
Receptor Profile	Expresses functional receptors for luteinizing hormone (LH), epidermal growth factor (EGF), androgens, estrogens, and progesterone, reflecting the physiological signaling competence of immature Leydig cells.
Growth Kinetics	It exhibits strong anchorage-dependent proliferation capability; under optimal conditions, the typical population doubling time is approximately 16 hours.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Remove and discard all spent complete culture medium from the culture dish. 2. Briefly rinse the cell monolayer with sterile PBS or a Ca2+/Mg2+-free 0.05% trypsin–0.53 mM (0.02%) EDTA solution to neutralize any residual serum containing active trypsin inhibitors. 3. Add 1.0 to 2.0 mL of trypsin-EDTA solution, ensuring the cell layer is completely covered. 4. Incubate at room temperature or 37°C for 1 to 3 minutes; monitor closely under an inverted microscope until the cells become rounded, loosen, and detach completely. 5. Immediately add 6.0 to 8.0 mL of pre-warmed complete culture medium to the dish to stop the enzymatic reaction. 6. Centrifuge the suspension at approximately 125 × g for 5 to 10 minutes. Decant the supernatant, gently resuspend the cell pellet in fresh complete culture medium, and distribute the cells into new vessels.
Thawing Protocol Note	TM3 cells are highly sensitive to excessively high culture medium alkalinity during the early recovery phase following thawing. Before adding the thawed cell suspension, the culture vessel containing fresh complete growth medium should be placed in a 5% CO2 incubator for at least 15 minutes to allow the medium's pH to reach the normal physiological range (7.0 to 7.6).
Medium Renewal	Every 3 to 4 days
Subcultivation Ratio	Split sub-confluent (70-80%) cultures at a standard ratio of 1:5 to 1:20.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete culture growth medium + 10% DMSO (or 50% Basal medium + 40% FBS + 10% DMSO)

Cat.No.	Product Name	Price
CSC-RO01226	Cas9 Stable Cell Line - TM3	Inquiry
CSC-RR00718	Luciferase Reporter Cell Line - TM3	Inquiry
CSC-RR00848	GFP Reporter Cell Line - TM3	Inquiry

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* For research use only. Not intended for any clinical use.

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