Germline-derived Pluripotent Stem Cells (gPS) | T2027

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Vector:

N/A

Species:

PND 35 Oct4-GFP Mice

Organ:

Testis

Passages:

N/A

Propagation:

Cells are grown on Mouse Embryonic Fibroblast Feeder Cells (Cat. No. T2001). The basal medium for this cell line is Prigrow III medium available at abm, Cat. No. TM003. To make the completed growth medium, add the following components to the base medium to the stated final concentration: 15% fetal bovine serum (TM999), MEM nonessential amino acids (ThermoFisher; 11140-050), 2 mM L-glutamine (G275), 50 µM mercaptoethanol (CH045), 10 ³ U/ml ESGRO, and Penicillin/Streptomycin (G255).
Carbon dioxide (CO₂): 5%, Temperature:37.0°C.

For in vitro glial differentiation: gPS cells aggregated to embryoid bodies. The base medium for this cell line is Prigrow III medium available from ABM (TM003). To make the completed growth medium, add the following components to the base medium to the stated final concentration: 15% fetal bovine serum (TM999), MEM nonessential amino acids (ThermoFisher; 11140-050), 2 mM L-glutamine (G275), 50 µM ?-mercaptoethanol (CH045), 10 ³ U/ml ESGRO (EMD Millipore; ESG1107), and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C.

Culture the cells with the complete growth media in presence of 10 ng/ml FGF2 (Z101455) and 20 ng/ml EGF (Z100135), followed by propagating cells in complete growth media with 10 ng/ml FGF2 (Z101455) and 10 ng/ml PDGF (Z100365) for 4 days. Differentiation is induced via 4-day growth factor withdrawal in presence of 30 ng/µl 3,3,5-tri-iodothyronine (T3; Sigma) and 200 µM ascorbic acid (AA; Sigma) in complete growth media. Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C.

For in vitro cardiomyocytes differentiation: gPS cells cultivated in hanging drops as aggregates to embryoid bodies for 48 hours. The base medium for this cell line is Prigrow III medium available from ABM (TM003). To make the completed growth medium, add the following components to the base medium to the stated final concentration: 20% fetal bovine serum (TM999), MEM nonessential amino acids (ThermoFisher; 11140-050), 2 mM L-glutamine (G275), 50 µM ?-mercaptoethanol (CH045), 10 ³ U/ml ESGRO (EMD Millipore; ESG1107), and Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 37.0°C.

After 48 hours, resuspend cells with 5 ml complete growth media and propagate cells as suspension in bacteriological petri dish for 5 days. Then plate embryoid bodies separately onto gelatin-coated 24 wells. Once attached, cardiomyocytes should appear between epithelial-like embryoid body cell outgrowth.

Concentration:

10⁶ cells/vial

Induction Method:

Germline-derived pluripotent stem cells (gPS) are generated from germline stem cells (GSCs) expressing Oct4-GFP promoter cultured under conversion culture conditions. The cell morphology and gene expression are comparable to the original derived GSC lines from the testes of PND 35 Oct4-GFP transgenic mice. These cells are pluripotent and can differentiate to different cell types such as cardiomyocytes and neural cells.

Pluripotency:

Positive for markers for alkaline phosphatase, and SSEA-1 by immunocytochemistry. It is capable of differentiating to cardiomyocytes or glial cells.

Growth Properties:

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Morphology:

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Markers:

N/A

Subculturing:

N/A

QC:

N/A

Safety:

BioSafety Level II

Shipping:

Dry Ice

Storage:

-180°C