GC-1 spg Cells

Name: CLS Cell Lines Service GmbH
Address: Dr.-Eckener-Straße 8, Eppelheim, 69214, DE
Telephone: +496221405780
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cryovial

Product number: 300375

Safety data sheet

Product sheet

Certificate of Analysis (CoA)

Description	The GC-1 spg cell line was immortalized through transfection with the pSV3-neo plasmid, which harbors the coding sequences for the SV40 large T antigen and neomycin resistance. This genetic modification not only provides resistance to certain antibiotics but also promotes the continuous growth of the cells by altering their cell cycle regulation, thus bypassing the Hayflick limit typical of primary cells. This process of immortalization allows the cells to maintain proliferative capacity while retaining key phenotypic characteristics of spermatogonia. Phenotypically, the GC-1 spg cell line exhibits characteristics that are indicative of a transitional stage between type B spermatogonia and primary spermatocytes, making it an especially relevant model for studying the early stages of spermatogenesis. The cells express two testis-specific isoproteins: cytochrome c and lactate dehydrogenase C4. These markers are crucial for studying cell metabolism and energy management during spermatogenesis, reflecting the unique metabolic pathways active in germ cells. The expression of these specific isoproteins underscores the cell line's utility in exploring the biochemical and physiological aspects of testicular cell function and development.
Organism	Mouse
Tissue	Testis
Applications	3D cell culture
Synonyms	GC-1spg, GC-1, GC1-SPG

Breed/Subspecies	BALB/c
Age	10 days
Gender	Male
Morphology	Epithelial
Cell type	Spermatocyte
Growth properties	Adherent

Citation	GC-1 spg (Cytion catalog number 300375)
Biosafety level	1
NCBI_TaxID	10090
CellosaurusAccession	CVCL_8872
GMO Status	GMO-S1: This murine testis cell line (GC-1 spg) contains an SV40 T-Antigen expression plasmid (pSV3neo) including a Tn5-neo resistance marker, supporting immortalization. The construct is stably integrated into mouse spermatogonial cells. This classification applies only within Germany and may differ elsewhere.

Viruses	Transformant: Simian virus 40 (SV40) T antigen

Culture Medium	DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Supplements	Supplement the medium with 10% FBS
Dissociation Reagent	Accutase
Subculturing	Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
Freeze medium	As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Thawing and Culturing Cells	Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
Incubation Atmosphere	37°C, 5% CO₂, humidified atmosphere.
Shipping Conditions	Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.
Storage Conditions	For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Sterility	Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.

Lot Number	Certificate Type	Date	Catalog Number
300375-050623	Certificate of Analysis	23. May. 2025	300375
300375-040825	Certificate of Analysis	22. Oct. 2025	300375

GC-1 spg Cells

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Certificate of Analysis (CoA)