Product Details

• Catalog Number: T0280
• Unit Size: 1 x 10⁶ cells / 1.0 ml
• Species: Human (Homo sapiens)
• Tissue: Brain (Fetal, 18 weeks gestation)
• Cell Type: Immortalized Astrocytes
• Growth Properties: Adherent, multipolar morphology
• Immortalization Method: Genetically modified using SV40 Large T antigen for extended proliferation.
• Expression Markers: NF-M, GDNF and NGF
• Biosafety Level: BSL-2
• Storage: Vapor phase of liquid nitrogen or below -130°C
• Shipping: Shipped on dry ice
• Format: Cryopreserved frozen cells

Overview

Immortalized Human Astrocytes (Fetal, SV40) serve as a stable and reproducible cellular model for investigating astrocyte physiology and function. These cells have been genetically immortalized with the SV40 Large T antigen, ensuring long-term proliferation while maintaining key astrocytic properties. They are ideal for research in neurodevelopment, neuroinflammation, and neurodegenerative diseases.

Key Features and Benefits

• Long-Term Viability: Genetically immortalized to enable extended culture durations.
• Consistent Performance: Provides reproducible results for multiple experimental applications.
• Neuroscience Applications: Ideal for studying astrocyte-neuron interactions, brain injury response, and neuroinflammatory processes.
• Optimized Culture Conditions: Designed to thrive in specialized media and extracellular matrices.
• Easy Handling: Adherent growth properties simplify maintenance in culture.

Culture & Handling Guidelines

Recommended Culture Conditions

• Coating: PriCoat™ ECM T25 Flasks (G999) or Applied Cell Extracellular Matrix (G422) are required for optimal cell adhesion.
• Growth Medium: PriGrow IV (TM004) supplemented with: 
  • 10% Fetal Bovine Serum (FBS)
  • 2 mM L-glutamine (G275)
  • 10 ng/mL recombinant human EGF (rhEGF, Z100139)
  • 1% Penicillin/Streptomycin Solution (G255)
• Incubation Conditions: 37°C, 5% CO₂ humidified atmosphere.
• Seeding Density: 40,000 – 50,000 cells/cm²
• Doubling Time: 49 – 59 hours

Thawing Protocol

1. Thaw cells rapidly in a 37°C water bath for no more than 2 minutes, keeping the vial cap above the water level to prevent contamination.
2. Decontaminate the vial exterior by spraying with 70% ethanol before transferring it into a biological safety cabinet.
3. Transfer the cell suspension into a 15 mL sterile conical tube containing 5 mL of pre-warmed complete growth medium.
4. Centrifuge at 125xg for 5-7 minutes.
5. Remove the supernatant without disturbing the cell pellet. Resuspend the pellet in fresh complete growth medium and seed into a pre-coated T25 flask.
6. Incubate under the recommended conditions and monitor cell recovery before passaging.

Subculturing Guidelines

1. Aspirate the spent medium and rinse cells with PBS.
2. Add 2-3 mL of pre-warmed 0.25% Trypsin-EDTA to the flask and incubate at 37°C until cells detach (2-10 minutes).
3. Neutralize Trypsin-EDTA by adding an equal volume of complete growth medium.
4. Transfer the suspension into a sterile centrifuge tube and centrifuge at 125xg for 5 minutes.
5. Remove the supernatant and gently resuspend the pellet in fresh complete growth medium.
6. Seed cells at the appropriate density into new culture vessels and incubate under recommended conditions.

Cryopreservation Guidelines

• Cryopreservation Medium: Complete growth medium supplemented with 10% DMSO or Cryopreservation Medium (TM024).
• Freezing Protocol: Freeze cells at a controlled rate of -1°C per minute before transferring them to liquid nitrogen storage.

STR Profiling (Short Tandem Repeat Analysis)

• D5S818: 11,13
• D13S317: 12,12
• TH01: 6,9.3
• AMEL: X,X

Related Products

• Recombinant Human TGFA – Z101935
• Recombinant Human FGF2 (E. coli) – Z101455
• Recombinant Human EGF – Z100139

Disclaimer

1. This product is intended for research use only. Not for human or animal therapeutic, diagnostic, or consumption purposes.
2. Growth parameters may vary based on user-specific culture conditions. It is recommended to validate in-house conditions before conducting critical experiments.
3. All sales are final. No warranty is provided for cell performance outside specified culture conditions.

References

1. Choi, Yong-Joon, et al. “PIN1 transcript variant 2 acts as a long non-coding RNA that controls the HIF-1-driven hypoxic response.” Scientific Reports 9.1 (2019): 10599.
2. Dozio, Vito, and Jean-Charles Sanchez. “Profiling the proteomic inflammatory state of human astrocytes using DIA mass spectrometry.” Journal of neuroinflammation 15 (2018): 1-14.