LLC-PK1 Cell Line

Brief Introduction
Product List

General Information
Organism	Sus scrofa, pig (porcine)
Cell Line Description	LLC-PK1 is a continuous porcine kidney epithelial cell line established by Robert N. Hull and his colleagues at Eli Lilly & Co. The cell line was derived from normal renal cortical tissue of a healthy, young (3-4 weeks old) male Hampshire pig. LLC-PK1 cells grow as an adherent monolayer and exhibit a highly polarized epithelial morphology. Globally, LLC-PK1 is recognized as the classic in vitro model of choice for studying the renal proximal tubule in the fields of renal physiology, pharmacology, and toxicology. These cells retain characteristics of differentiated epithelial cells, including the formation of apical microvilli, functional tight junctions, and active sodium-dependent transport systems. The cell line is widely used as a standard model for investigating transepithelial transport, drug-induced nephrotoxicity, and host-pathogen interactions specific to pigs.
Tissue	Kidney
Cell Type	Epithelial cell
Breed / Strain	Hampshire
Disease	Normal / Non-malignant tissue origin (Spontaneously immortalized)
Morphology	Epithelial-like; uniform polygonal cells growing in tightly packed, cobblestone monolayers
Gender	Male
Age	3 to 4 weeks
Product Format	Frozen
Growth Mode	Adherent
Biosafety Level	1 (Biosafety classification matches standard laboratory parameters for non-infectious agricultural biomaterials)
Applications	1. Highly cited in vitro model for studying renal proximal tubule transport mechanisms (e.g., coupled transport of glucose, amino acids, sodium, and phosphate); 2. Standard preclinical target cell line for screening drug-induced nephrotoxicity and cell membrane damage (using MTT or LDH release assays); 3. Model for simulating cellular mechanisms of apoptosis and oxidative stress pathways in renal cells following chemical-induced injury; 4. Host cells for studying porcine enteric pathogenic coronaviruses; 5. Model for investigating intracellular protein sorting pathways, apical/basolateral membrane polarization, and tight junction barrier permeability.
Shipped In	Dry ice
Storage Temperature	−196°C
Characteristics
Tumorigenic	Yes; can form poorly differentiated carcinomas when inoculated into immune-deficient or newborn rodents, despite its normal embryonic/young tissue origin.
Transformant	None; immortalized via continuous spontaneous cell replication adaptation during serial passage selections.
Karyotype	Aneuploid/near-diploid; the modal chromosome number is typically around 38. However, structural rearrangements, deletions, and marker chromosomes are present.
Physiological Markers	They highly express proximal tubule-associated enzymes, including alkaline phosphatase, γ-glutamyl transpeptidase (γ-GTP), and leucine aminopeptidase; and possess active sodium-coupled transporters for hexoses (α-methyl-D-glucoside) and amino acids.
Growth Kinetics	Robust and rapid adherent proliferation; population doubling time spans approximately 24 to 26 hours.
Mycoplasma Test	Negative
Culture Conditions and Handling
Subculturing	1. Completely remove and discard all spent complete culture medium from the culture flask. 2. Briefly rinse the cell monolayer with sterile, Ca2+/Mg2+-free PBS buffer to remove residual serum containing trypsin inhibitors. 3. Add 1.0 to 2.0 mL of 0.25% trypsin-0.53 mM (0.03%) EDTA solution, ensuring the cell layer is completely submerged. 4. Incubate at 37°C for 3 to 8 minutes; monitor under an inverted microscope until the cells become rounded, loosen, and fully detach. Key measure to prevent clumping: Do not tap, shake, or vigorously agitate the flask while waiting for the cells to detach. If cells are difficult to detach, continue incubation at 37°C to facilitate dissociation. 5. Add 6.0 to 8.0 mL of pre-warmed complete culture medium to the flask and gently pipette up and down to resuspend the cells until a uniform single-cell suspension is obtained. 6. Centrifuge at approximately 125 × g for 5 to 7 minutes, discard the supernatant, gently resuspend the cell pellet in fresh complete culture medium, and distribute the suspension into new culture vessels.
Thawing Protocol Note	During the early stages of thawing and recovery, LLC-PK1 cells are highly sensitive to sudden changes in the alkalinity of the culture medium. Before adding the thawed cell suspension, the culture vessel containing fresh complete 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 times per week
Subcultivation Ratio	Split optimal, confluent cultures at a standard ratio of 1:3 to 1:8.
Culture Conditions	Atmosphere: Air, 95%; CO2, 5%; Temperature: 37°C
Cryopreservation	90% Complete culture medium + 10% DMSO (or 95% complete culture medium + 5% DMSO)

Cat.No.	Product Name	Price
CSC-RO02570	Cas9 Stable Cell Line - LLC-PK1	Inquiry
CSC-RR00884	GFP Reporter Cell Line - LLC-PK1	Inquiry
CSC-RR00933	Luciferase Reporter Cell Line - LLC-PK1	Inquiry

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

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