TM4 Cell Line

Brief Introduction
Product List

General Information
Organism	Mus musculus, mouse
Cell Line Description	TM4 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 Sertoli cells, isolated from the normal testes of 11- to 13-day-old male BALB/c mice (specifically of the BALB/c nu/+ genotype). TM4 cells exhibit an epithelial-like morphology and are anchorage-dependent, growing as a tightly adherent monolayer. In the fields of reproductive biology and endocrinology, TM4 is recognized as the "gold standard" non-tumorigenic in vitro model for studying immature Sertoli cells. The cell line retains various functional characteristics of juvenile Sertoli cells, including responsiveness to specific hormones and associated metabolic biosynthetic pathways. It is widely used as a standard model for investigating the mechanisms supporting spermatogenesis, the effects of endocrine-disrupting chemicals, and the dynamics of the blood-testis barrier.
Tissue	Testis; Seminiferous tubules / Sertoli cells
Cell Type	Sertoli cell
Strain	BALB/c
Disease	Normal / Non-malignant tissue origin
Morphology	Epithelial-like
Gender	Male
Age	11 to 13 days
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 (Biosafety classification aligns with global standard laboratory parameters for non-infectious mouse-derived biomaterials)
Applications	1. Highly cited in vitro model for studying endocrine, paracrine, and autocrine regulation of Sertoli cell function; 2. Standard target substrate for modeling reproductive toxicology, heavy metal insult, and environmental endocrine-disruptor exposure; 3. Investigating spermatogenic support mechanisms, cell-to-cell communication, and junctional protein dynamics of the blood-testis barrier; 4. Dissecting hormonal pathways of follicle-stimulating hormone (FSH), androgens, and thyroid hormones in testicular cells; 5. Host system for studying cellular transport, retinol metabolism, and lipid-associated signaling cascades.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	No, non-tumorigenic; repeatedly verified to not form tumors when subcutaneously or intratesticularly engrafted into BALB/c-$nu/nu$ (nude) mice.
Transformant	None; spontaneously immortalized during extended serial cultivation and selective expansion.
Hormone Responsiveness	Responds to purified follicle-stimulating hormone (FSH) with an increase in cyclic AMP (cAMP) production, but exhibits no response to luteinizing hormone (LH).
Secretory & Synthesis Profile	1. Secretions: Capable of synthesizing and secreting a unique retinol-binding protein (RBP), transferrin, and H-Y antigen. 2. Plasminogen activator: Basal production of plasminogen activator is low, but it is significantly stimulated by FSH and even more strongly stimulated by retinoic acid.
Receptor Profile	The endogenous expression of functional androgen, estrogen, and progesterone receptors reflects the physiological signaling capacity of immature testicular Sertoli cells.
Growth Kinetics	Robust adherent proliferation; typical population doubling time is approximately 18 hours under optimal conditions.
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 protease 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 2 to 5 minutes; monitor closely under an inverted microscope until the cells become rounded, loosen, and fully detach. 5. Immediately add 6.0 to 8.0 mL of pre-warmed complete culture medium to the dish and gently pipette up and down to resuspend the cells and achieve a uniform single-cell suspension. 6. Centrifuge at approximately 125 × g for 5 to 10 minutes, discard the supernatant, gently resuspend the cell pellet in fresh complete culture medium, and distribute into new vessels.
Thawing Protocol Note	During the early recovery phase after thawing, TM4 cells are highly sensitive to excessively high alkalinity in the culture medium. 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	2 to 3 times per week
Subcultivation Ratio	Split sub-confluent (70–80%) cultures at a standard ratio of 1:10 to 1:20, or seed at a density of approximately 1.0 X 10^4 cells/cm2. Cells resuscitated from frozen stock should be split at a lower ratio of 1:3 at the first passage.
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-RR00849	Luciferase Reporter Cell Line - TM4	Inquiry
CSC-RR00850	GFP Reporter Cell Line - TM4	Inquiry
CSC-RO02586	Cas9 Stable Cell Line - TM4	Inquiry

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

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