Immortalized Rat Hippocampal Cells (H19-7)

Immortalized Rat Hippocampal Cells (H19-7) are derived from hippocampi that were dissected from E17 Holtzman rat embryos and subsequently immortalized via retroviral infection with tsA58 and U19tsa temperature sensitive mutants of the simian virus 40 large tumor antigen gene. These cells exhibit polygonal morphology prior to differentiation, after which they became elongated with thickened cell bodies and multiple extended processes. Differentiation of H19-7 cells depends upon the inactivation of the ts large tumor antigen and the environmental conditions. It was found that H19-7 cease DNA replication and cell division upon differentiation and are responsive to factors secreted by primary glia. H19-7 cells are useful tools in analyzing the development and regulation of hippocampal trophic interactions, an extremely interesting area given that the hippocampal formation expresses trophic signals that support the development and maintenance of synaptic inputs from the septal region, which in turn are invaluable for normal cognition and are particularly vulnerable in diseases such as Alzheimer's.

Immortalization Method: Immortalized via retroviral transduction of the tsA58 and U19tsa simian virus 40 large tumor antigen.

BioSafety Level: II

Species: Rat

Source Organ: Hippocampus

Growth Properties: Adherent

Morphology: Elongated

Population Doubling: 22 - 31 hours

Seeding Density: N/A

Markers: NFP, MAP-2, GAP-43 post-differentiation

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow III medium, available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, L-glutamine (G275), 0.2 mg/ml G418, and Penicillin/Streptomycin solution(G255) to a final concentration of 1%.
Change media every 2-3 days.
Carbon dioxide (CO₂): 5%, Temperature: 33.0°C.

Protocol and Media for Differentiation:
The base medium for this cell line is Prigrow III medium, available at abm, Cat. No. TM003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 1%, 20 nM hydrocortisone, 0.3 nM triiodothyronine, 0.1 mM putrescine, 20 nM progesterone, 1 pM estradiol, 30 nM sodium selenite, transferrin at 1 μg/ml, and insulin at 5 μg/ml. On Day 3, add 10 nM phorbol 12,13-dibutyrate (PBt2), the differentiation agent, to the culture.
Carbon dioxide (CO₂): 5%, Temperature: 39.0°C (nonpermissive).

* Do not use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Immunostaining

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C