|Description||The 15P-1 cell line was derived from transgenic mouse testicular cells that express PyLT in the seminiferous epithelium and show characteristics of Sertoli cells, such as the transcription of the WT1 and Steel genes. The 15P-1 cells exhibit phagocytic activity, which is increased when interacting with germ cells. The 15P-1 cells also aid in the differentiation of diploid premeiotic germ cells into haploid spermatids expressing the Prm-1 gene. When the 15P-1 cells are co-cultured with immature testicular cells from 9-day-old animals, the cells form tetrads of haploid cells with the morphology of round spermatids and initiate protamine transcription. Also, the 15P-1 cells release a material that is sensitive to proteases and that shows broad-spectrum antibacterial activity. The activity of this material is increased by fractions that are rich in round spermatids and nerve growth factors. Both Sertoli cells and 15P-1 cells express the KL protein, which is a ligand of the Kit receptor.|
Identifiers / Biosafety / Citation
|Citation||15P-1 (CLS catalog number 305191)|
Expression / Mutation / Susceptibility
|Medium supplements||10% FBS, 4.0 mM L-glutamine, 4.5 g/L glucose, 3.7 g/L NaHCO3|
|Subculturing||Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium.|
|Split ratio||1:2 to 1:5|
|Fluid renewal||2 to 3 times per week|
|Freeze medium||CM-1 (CLS catalog number 800100) or CM-ACF (CLS catalog number 806100)|
|Handling of cryopreserved cultures||The cells come deep-frozen shipped on dry ice. Please make sure that the vial is still frozen. If immediate culturing is not intended, the cryovial must be stored below -150 degree Celsius after arrival. If immediate culturing is intended, please follow the below instructions: Quickly thaw by rapid agitation in a 32 degree Celsius water bath within 40-60 seconds. The water bath should have clean water containing an antimicrobial agent. As soon as the sample has thawed, remove the cryovial from the water bath. A small ice clump should still remain and the vial should still be cold. From now on, all operations should be carried out under aseptic conditions. Transfer the cryovial to a sterile flow cabinet and wipe with 70% alcohol. Carefully open the vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of culture medium (room temperature). Resuspend the cells carefully. Centrifuge at 300 x g for 3 min and discard the supernatant. The centrifugation step may be omitted, but in this case the remains of the freeze medium have to be removed 24 hours later. Resuspend the cells carefully in 10 ml fresh cell culture medium and transfer them into two T25 cell culture flasks. All further steps are described in the subculture section.|
|Handling of proliferating cultures||One or two cell culture flasks come filled with cell culture medium. Collect the entire medium in 1 or 2 x 50 ml centrifuge tubes, respectively. Carefully add 5 ml of cell culture medium to each T25 cell culture flask. Control the cell morphology and confluency under the microscope. Incubate at 32 degree Celsius for a minimum of 24 hours. Spin down the collected medium at 300 x g for 3 minutes to collect the cells which may have detached during transit. If a cell pellet is visible, resuspend the cells in 5 ml of cell culture medium and transfer to a T25 cell culture flask. Incubate at 32 degree Celsius for a minimum of 24 hours.|
Quality control / Genetic profile / HLA
|Sterility||Mycoplasma contamination was excluded through PCR-based and luminescence-based mycoplasma assays. Bacterial or fungal contaminations are detected through daily visual cell monitoring.|
To identify mycoplasma contaminations we perform PCR-based and luminescence-based mycoplasma assays. We further determine any bacterial or fungal contamination through our standardized manufacturing processes.
Besides genomic DNA, RNA, cell pellets, and cell lysates in bulk, we can offer large quantities of assay-ready cells, plated cells in multiple formats, and frozen or growing cells. Contact us to receive a quote.
Each manufactured batch of cell lines* is authenticated via STR analysis. Contact us if you require a publication-ready STR report for your cells (*human, hamster, mouse, rat, and dog cells).
HLA characterization is available from more than 200 cell lines. HLA class I -A, B, C, and Class II HLA-DPA1, -DPB1, -DQA1, DQB1, and DRB1 alleles were obtained by next-generation sequencing methodologies (NGS) for class I and class II alleles.