Pricella® Primary Liver-Derived Cell Isolation Kits: Achieve High-Purity Liver-Derived Cell with Ease

Isolating and culturing primary cells is a cornerstone of life science research, providing a more accurate representation of the physiological state of cells in vivo. To help researchers overcome the technical challenges of primary cell isolation, Pricella® has developed a series of high-efficiency, easy-to-use isolation kits covering six major tissue types: brain, vascular tissue, liver, heart, bone marrow, and heart. These kits are designed to deliver high-purity, high-viability cells quickly and consistently.

In this edition of Cell Culture Academy, we spotlight the Primary Liver-Derived Cell Isolation Kits , detailing their key features, typical applications, and practical tips.

Ⅰ. Primary Liver-Derived Cell Types and Their Research

The liver is a complex organ composed of multiple functional cell types that work together to regulate metabolism, mediate immune responses, and support tissue repair. In research, liver cells are generally classified into two main categories: hepatic parenchymal and non-parenchymal cells.

1.Hepatic Parenchymal Cells 

Hepatic parenchymal cells are the liver’s primary metabolic cells and form the core of the hepatic parenchyma. They are relatively large, typically round, and some exhibit distinct binucleation. Hepatic parenchymal cells carry out essential liver functions, including metabolism, detoxification, and bile production. As such, they are a primary focus in studies of liver pathophysiology, such as metabolic disorders and fatty liver disease.

2. Hepatic Non-parenchymal Cells

Non-parenchymal liver cells do not directly perform the liver’s core metabolic functions, but they play key roles in maintaining metabolic homeostasis through immune regulation, structural support, and microenvironment maintenance. They are generally divided into three main types:

A.Hepatic Stellate Cells (HSCs): Accounting for roughly 30% of non-parenchymal cells, HSCs reside in the space of Disse, between liver sinusoids and Hepatic Parenchymal Cells . In a healthy liver, they remain quiescent and primarily store vitamin A. When the liver is injured or exposed to inflammatory stimuli, HSCs become activated, adopting a myofibroblast-like phenotype and contributing to tissue repair and fibrosis regulation. Research on HSCs often focuses on liver fibrosis and the regenerative liver microenvironment.

B.Kupffer Cells: These resident liver macrophages are located mainly within the liver sinusoids. They phagocytose foreign particles, clear senescent cells and pathogens, and mediate immune responses. Key research areas include infection-related immunity, the inflammation-to-fibrosis transition, and other liver diseases.

C.Liver Sinusoidal Endothelial Cells (LSECs): Characterized by fenestrations and the absence of a complete basement membrane, LSECs form a highly permeable barrier between blood and hepatic parenchymal cells. They regulate the exchange and transmembrane transport of substances such as oxygen, hormones, and metabolites, playing a vital role in maintaining hepatic microcirculation and tissue homeostasis. Research on LSECs primarily addresses sinusoidal microcirculation disorders, liver disease microenvironment regulation, and mechanisms of transmembrane substance transport.

Ⅱ. Pricella® Primary Liver-Derived Cell Isolation Kits

1. Product Catalog

2. Kit Components and Brief Description of Their Functions

Washing Solution: Used for rinsing liver tissues while effectively preserving tissue and cell viability. It also helps minimize the risk of contamination.

Specific Separation Solution: Based on differences in cell density, cells are separated by density gradient centrifugation, which distributed them into distinct layers within the centrifuge tube, thereby achieving separation and purification.

Digestive Enzyme & Diluent of Digestive Enzyme: Prepared at the appropriate ratio, and used to enzymatically digest extracellular matrix proteins, yielding a single-cell suspension.

Basic Culture Medium and Supplement: Provide essential nutrients to support cell growth and promote the preferential expansion of target cell types.

Cell Filter: Removes undigested tissue fragments and debris through physical filtration, improving the purity of the resulting cell suspension.

Pre-perfusion Solution (Hepatic Parenchymal Cells): Used to remove blood and inhibit Ca2+-and Mg2+-dependent adhesion, preparing the liver for subsequent perfusion.

3. Cell Images from Kit Isolation (Partial)

	Rat Hepatic Stellate Cells (α-SMA)	Rat Kupffer Cells (CD68)	Rat Liver Sinusoidal Endothelial Cells (CD31)	Mouse Hepatic Parenchymal Cells (CK-18)
Microscopic View of Cell Morphology
Immunofluorescence Identification

Primary Liver-Derivedcells isolated using Pricella® kits exhibit excellent experimental characteristics: they show strong adherence, typical cellular morphology, and stable physiological conditions. Target cell-specific markers are robustly expressed, with purity exceeding 90%. These cells demonstrate high reproducibility and consistent results, making them well-suited for downstream experimental applications.

Ⅲ . Common Issues in Primary Liver-Derived Cell Isolation and Culture

1.What to do if hepatocyte perfusion is difficult or cell viability is low?

Perform mock perfusion with PBS in normal rats. Proceed to the actual experiment only after successfully completing at least two mock runs.

2.What should be considered when using separation solution to isolate and purify liver cells?

Ensure the cell suspension and the underlying separation solution form a clear layer. If layering is unclear, first gently overlay a small volume using a low-volume pipette, then use a larger-volume pipette or syringe if necessary.

3.Are there age restrictions for experimental mice?

Different liver cell types have varying sensitivities to the age of the animals. It is recommended to strictly follow the age and number of animals specified in the kit instructions.

A. Liver sinusoidal endothelial cell and Kupffer cell isolation and culture kits are suitable for 2-12-day-old mice or rats. If the amount of liver tissue obtained is insufficient, the number of animals can be increased.

B. Hepatic stellate cell isolation and culture kits are suitable for 1-2-day-old neonatal rats.

C. Hepatic parenchymal cells isolation and culture kits are recommended for 4-12-week-old mice and 2-8-week-old rats.

4.Hepatic parenchymal cells perfusion is consistently suboptimal—how can this be addressed?

If the perfusion rate is correct but the perfusion outcome is poor, it is often due to errors in needle insertion or fixation. It is recommended to practice needle placement and perfusion techniques to improve results. 

5.Mixed contamination of liver sinusoidal endothelial cells and Kupffer cells—how can this be resolved?

These two cell types have similar isolation procedures and are prone to cross-contamination. Purification can be achieved using differential adhesion and differential digestion, taking advantage of the fact that Kupffer cells attach more quickly, are more resistant to digestion, and have lower nutritional requirements than liver sinusoidal endothelial cells. 

6.What causes incomplete or over-digestion of liver sinusoidal endothelial cells and Kupffer cells, and what are the consequences?

Incomplete digestion may result from using too much tissue, insufficient mincing, or limited enzyme contact due to upright centrifuge tubes.

Over-digestion can occur if low-temperature digestion is prolonged. 

Both scenarios can negatively affect cell yield and purity, so it is essential to strictly follow the kit instructions. 

7.Can liver-derived cells be passaged? What is the maximum number of passages? 

The passage potential varies among different liver cell types: 

A.Hepatic parenchymal cells cannot be digested or passaged; the use of any digestive enzymes is strictly prohibited. 

B.Hepatic stellate cells can be digested with trypsin and passaged in vitro for approximately three passages. 

C.Liver sinusoidal endothelial cells can be digested with trypsin; rat-derived cells can be passaged 2-3 times, while mouse-derived cells can be passaged only 1-2 times. 

D.Kupffer cells are largely non-proliferative. Trypsin digestion is difficult (requires more than 5 min), and prolonged digestion significantly affects cell viability. Passaging is not recommended. 

8.Why are there no cells in the target layer after separation with the separation solution? 

The absence of cells in the target layer may result from insufficient tissue digestion, low cell numbers, or a high proportion of dead cells, indicating a failed experiment. 

9.How long can different liver cells be cultured? 

A.Hepatic parenchymal cells typically display their characteristic morphology on the first day but begin to dedifferentiate after day 2. They can be cultured for more than 10 days, but it is recommended to complete experiments within 3 d.

B. Kupffer cells and liver sinusoidal endothelial cells can generally be cultured for 1-2 weeks.

C. Hepatic stellate cells can be cultured for approximately 2 weeks.