Immortalized Drosophila Embryonic Cells (R3)

The immortalized Drosophila Embryonic Cells (R3) was established by serial passaging of primary cultures from Act5C > UAS-Ras^V12 embryos. The Ras^V12 immortalized cell lines have significantly up-regulated cell-cycle and cell-division gene expression including gene targets of the E2 promoter binding factor/retinoblastoma protein (E2F/RB) pathway. Specifically, R3 express upregulated Cyc A, tum, sti, pav, stg, and dup. The cell lines are in a proliferative undifferentiated progenitor-like state associated with increasing levels of Polycomb Group (PcG) transcripts during the immortalization process. Related to adult muscle precursors (AMPs), a stem cell-like population contributing to adult muscles and sharing properties with vertebrate satellite cells, the immortalized R3 cell line retained the capacity for myogenic differentiation when treated with the steroid hormone ecdysone. The cell line is recommended as a tool for Drosophila embryogenesis and muscle cell differentiation and proliferation.

Immortalization Method: Serial passaging of primary cultures from Act5C > UAS-Ras^V12 embryos

BioSafety Level: II

Species: Insect

Source Organ: Embryos

Growth Properties: Adherent

Morphology: Spindle shaped

Population Doubling: N/A

Seeding Density: Thaw entire contents into an appropriate T25 flask as specified in the Propagation instructions.

Markers: N/A

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Schneider's Insect Medium (TM034). To make the completed growth medium, add the following components to the base medium: heat-inactivated fetal bovine serum (TM999)* to a final concentration of 10%, and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%.
Carbon dioxide (CO₂): 5%, Temperature: 22.0°C.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: (1) Inducing muscle cell differentiation by treating cells with ecdysone and perform immunofluorescence staining and qPCR to assess the upregulated expression of mhc and Tropomyosin (Tm).
(2) Up-regulated genes in DPCs are analyzed by DAVID (the Database for Annotation, Visualization and Integrated Discovery).

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C