HLE-B3 Cell Line

Brief Introduction
Product List

General Information
Organism	Homo sapiens, human
Cell Line Description	HLE-B3 (officially registered as B-3 in major cell banks) is an immortalized human lens epithelial cell line established in 1994 by U.P. Andley, T.P. Fleming, and their colleagues at the Washington University School of Medicine. The cell line was derived from normal lens epithelial tissue obtained from an infant (aged 5 to 12 months) undergoing clinical treatment for retinopathy of prematurity. Immortalization was achieved by infecting primary lens epithelial cells at passage 3 (60% confluence) with the hybrid virus Ad12-SV40. HLE-B3 cells exhibit an epithelial morphology and grow as a tightly adherent monolayer. In the fields of ophthalmology and toxicology, HLE-B3 serves as a global standard in vitro model for studying human lens cell biology and the pathogenesis of cataracts. Retaining the native ability to synthesize lens-specific proteins (such as crystallins), this cell line offers an indispensable alternative to primary human lens tissue.
Tissue	Eye; derived from ocular lens epithelium
Cell Type	Lens Epithelial Cell (LEC)
Disease	Normal / Non-malignant tissue origin (Immortalized engineered line)
Morphology	Epithelial-like
Age	Infant
Product Format	Frozen
Growth Mode	Adherent Monolayer
Biosafety Level	1 (Biosafety classification aligns with standard laboratory safety parameters for non-infectious, replication-incompetent virus-transformed human lines; cells cease producing infectious viral particles after a few passages).
Applications	1. Directly simulate human cataract formation, lens epithelial cell senescence, and age-related ocular degenerative changes in vitro; 2. Serve as a primary target cell line for ophthalmic toxicology screening to evaluate the safety, toxicity, and efficacy of anti-cataract drugs; 3. Elucidate cellular pathways under oxidative stress, particularly hydrogen peroxide (H2O2)-induced damage and thioltransferase-mediated repair mechanisms; 4. Investigate the epithelial-mesenchymal transition (EMT) process in lens cells, which mimics the pathological changes associated with posterior capsule opacification (PCO) or "secondary cataract"; 5. Assess the effects of ultraviolet radiation, metabolic stress (such as factors associated with diabetic cataracts), and calcium channel blockade on lens cell proliferation.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	No, non-tumorigenic under standard physiological baselines.
Transformant	Adenovirus 12-SV40 (Ad12-SV40) hybrid virus. Expresses SV40 large T-antigen.
Expression Profile	Human β-crystallins and γ-crystallins were synthesized and expressed, and their expression was monitored and verified by Western blot analysis.
Growth Kinetics & Lifespan	Highly proliferative; can be continuously maintained in culture for over 76 population doublings without any detectable decrease in proliferative capability. Typical population doubling time is approximately 24 to 36 hours.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Completely remove and discard the old complete culture medium from the culture vessel. 2. Note on trypsin sensitivity: HLE-B3 cells are extremely sensitive to trypsin-induced damage. 3. Briefly rinse the cell monolayer with 0.25% trypsin-0.53 mM EDTA solution at room temperature (for less than 30 seconds) to thoroughly remove residual serum components containing trypsin inhibitors; discard the rinse solution immediately. 4. Add fresh trypsin-EDTA solution (approximately 1.0 to 2.0 mL for a T-25 flask) to cover the cell layer. 5. Incubate at room temperature for 60 seconds or less. Observe under an inverted microscope until the cells round up and detach. Do not incubate at 37°C or prolong the incubation time, as this will reduce cell viability. 6. Add 6.0 to 8.0 mL of pre-warmed complete growth medium to rapidly stop the enzymatic reaction. 7. Gently pipette up and down to disperse the cells into a single-cell suspension, centrifuge at approximately 125 × g for 5 minutes, carefully discard the supernatant, gently resuspend the cells in fresh complete growth medium, and distribute them into new culture vessels.
Thawing Protocol Note	HLE-B3 cells are highly sensitive to sudden pH changes during the thawing process. Before adding the thawed cell suspension, the culture vessel containing fresh complete growth medium should be placed in a 5% CO2 incubator for at least 15 minutes to allow the medium's pH to reach and equilibrate within the normal physiological range (7.0 to 7.6).
Medium Renewal	2 to 3 times per week
Subcultivation Ratio	Split confluent cultures (80% density) at a standard ratio of 1:2 to 1:4.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete culture growth medium + 10% DMSO (or 70% Complete culture medium + 20% FBS + 10% DMSO)

Cat.No.	Product Name	Price
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CSC-RR00908	GFP Reporter Cell Line - HLE-B3	Inquiry
CSC-RR00954	Luciferase Reporter Cell Line - HLE-B3	Inquiry

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* For research use only. Not intended for any clinical use.

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