In the field of cellular biology research, the HFL1 cell line, as a classic human embryonic lung fibroblast model, is widely utilized in virology, vaccine development, drug screening, and studies on aging mechanisms. However, traditional HFL1 cells are a finite-passage cell line, and excessive passaging can lead to cellular senescence, altered growth characteristics, and even compromise the reliability and reproducibility of experimental results. To ensure data accuracy, it is generally recommended to complete experiments within 3 passages after cell arrival, which inevitably increases both time and economic costs.

Hycyte has successfully developed the HFL1/TERT-SV40 cell line using advanced dual-transformation technology incorporating SV40T antigen gene and hTERT (telomerase reverse transcriptase), with G418 resistance. This cell line not only retains the classic properties of HFL1 cells but also exhibits excellent passaging capability.

Cell Name: TERT-SV40 Transformed Human Embryonic Lung Fibroblasts
Cell Synonyms: HFL1/TERT-SV40
Catalogue No.: TCH-C621
Species: Human
Tissue Origin: Lung
Disease Characteristics: /

Morphology: Fibroblast-like
Growth Properties: Adherent
Culture Medium: F12K + 10% FBS + 1% P/S
Matching Medium Catalog Number: TCH-G621
Passage Ratio: 1:2, with medium change every 2-3 days
Doubling Time: 24-48 h
Culture Conditions: Atmosphere: 95% air + 5% CO₂; Temperature: 37°C
Freezing Conditions:: 60% basal medium + 30% FBS + 10% DMSO, stored in liquid nitrogen
Recommended Starfish HyCyte® One-Step Cryopreservation Solution (Ready-to-use, serum-free, no programmed freezing required): No. GUCP-R201
Quality Control: Negative for bacteria, fungi, and mycoplasma

Culture Notes:

1. Passaging Capacity of HFL1/TERT-SV40:

   • ① Cell status is sensitive to medium changes. Use Hycyte complete medium (No. TCH-G621) to avoid compromising cell health and passaging ability.

   • ② When using Starfish’s complete medium and passaging at 1:2, the cells are guaranteed to undergo at least 20 additional passages post-delivery.

2. Digestion of HFL1/TERT-SV40:

   • ① These cells are sensitive to trypsin. Use Trypsin-EDTA (0.25%) with phenol red (No. GUTP-R002) for digestion at 37°C for 1–2 minutes. Digestion time may vary slightly depending on the brand of trypsin and cell density. Stop digestion when 80% of cells round up and detach; over-digestion may adversely affect cell viability.

   • ② A small number of floating cells may appear the day after passaging. This is normal and can be removed via medium change without affecting cell culture.