Immortalized Fibroblast Cryopreservation Medium

Catalog Number: IM03 
Product Format: Frozen
Product Volume: 20 mL
Product Usage: Research Use Only. Not for Clinical Use.

Custom Options

• DMSO-free (for sensitive applications)

• Serum-containing (for primary fibroblasts)

Product Description

A specialized, serum-free cryopreservation medium designed for immortalized fibroblast cell lines (e.g., NIH/3T3, MRC-5, hTERT-BJ1). Formulated to maintain ≥95% post-thaw viability while preserving morphology, proliferation potential, and biomarker expression.

Key Features

✔ High recovery rates (≥95% viability for most fibroblast lines)
✔ Ready-to-use (no preparation required)
✔ Xeno-free and animal component-free formulation
✔ Validated for:

• Skin fibroblasts (e.g., BJ-5ta)

• Lung fibroblasts (e.g., MRC-5)

• Tumor-associated fibroblasts (e.g., CAFs)

✔ Compatible with:

• Controlled-rate freezers

• Mr. Frosty™ passive freezing containers

• Liquid nitrogen vapor phase storage

Physicochemical Properties

• pH: 7.2–7.4

• Osmolality: 1000–1200 mOsm/kg

• Sterility: 0.22 µm filtered

Recommended Protocol

A. Freezing Procedure

1. Harvest cells at 80–90% confluency using 0.25% Trypsin-EDTA.

2. Neutralize trypsin, pellet cells (200 × g, 5 min).

3. Resuspend in growth medium at 2–5 × 10⁶ cells/mL.

4. Mix 1:1 with chilled cryopreservation medium (final cell density: 1–2.5 × 10⁶ cells/mL).

5. Aliquot 1 mL per cryovial and freeze:

   • Programmable freezer: –1°C/min to –80°C, then transfer to liquid nitrogen

   • Passive cooling: Mr. Frosty™ at –80°C overnight, then transfer to liquid nitrogen

B. Thawing Procedure

1. Quick-thaw in a 37°C water bath (≤90 seconds)

2. Dilute 1:10 in pre-warmed growth medium

3. Centrifuge at 200 × g for 5 minutes to remove cryoprotectants

4. Reseed at 2× normal density for recovery passage

Quality Control – Performance Data (NIH/3T3)

Safety Testing

• Sterility: 14-day incubation – no microbial growth

• Mycoplasma: PCR-negative

• Endotoxin: <0.1 EU/mL

Storage & Stability

• Unopened: 12 months at 2–8°C

• Opened: 4 weeks at 2–8°C

• Note: Do not freeze the cryopreservation medium itself

Troubleshooting Guide

Ordering Information

References

1. Cryobiology (2022): “Optimized Cryopreservation of Fibroblasts”

2. ATCC® Guidelines: NIH/3T3 Cryopreservation Protocol

Product Usage

Cells are offered for Research Use Only. Not for Clinical Use.