Pricella® Primary Cardiac-Derived Cell Isolation Kits: Say Goodbye to Cardiac-Derived Cell Isolation Challenges

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, cartilage, 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 Cardiac-Derived Cell Isolation Kits, detailing their key features, typical applications, and practical tips.

Ⅰ. Primary Cardiac-Derived Cell Types and Their Research

Non-immune cells in the heart not only provide structural support and sustain blood-pumping and vascular functions but also play crucial roles (changed from critical roles → crucial roles is slightly more natural in this context, though critical is not wrong) in processes such as injury repair and inflammation regulation. The three main types commonly studied are:

1.Cardiomyocytes

These are the primary contractile cells of the heart, capable of spontaneous beating and typically irregularly star-shaped (changed from irregular star-like in shape for smoother expression). Their key properties, excitability, autorhythmicity, conductivity, and contractility, enable continuous blood circulation and fluid regulation. However, cardiomyocytes are terminally differentiated with extremely limited proliferative capacity. Once lost due to ischemia, hypoxia, or injury (e.g., myocardial infarction), they cannot be effectively replaced through self-regeneration, contributing to the poor prognosis of many heart diseases.

2.Cardiac Fibroblasts

As the most abundant interstitial cells, cardiac fibroblasts are distributed among cardiomyocytes and provide structural support by secreting extracellular matrix components such as collagen. Following cardiac injury, they proliferate to form scar tissue. While this response is essential for repair, excessive activation can lead to myocardial fibrosis, making fibroblasts a double-edged sword in both recovery and pathological remodeling.

3.Cardiac Microvascular Endothelial Cells

These cells form a typical “cobblestone” arrangement between circulating blood and myocardial interstitial fluid, acting as a barrier for material exchange. Through specialized intercellular junctions, they regulate vascular permeability. Under normal conditions, they contribute to key processes including vasodilation, leukocyte recruitment, inflammation, and angiogenesis-mediated by nitric oxide, cytokines, and junctional proteins. Structural or functional abnormalities in these cells are central to the pathology of cardiovascular diseases such as ischemic heart disease and myocarditis.

Ⅱ. Pricella® Primary Cardiac-Derived Cell Isolation Kits

1. Product Catalog

2. Kit Components and Brief Description of Their Functions

Washing Solution: Used for rinsing heart tissues, provides a liquid environment for tissues and cells, and serves to protect and inhibit bacterial growth.

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

Digestive Solution: 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.

Screening Solution: By selectively inhibiting the growth of nontarget cells, the purity of target cells can be further improved.

3. Cell Images from Kit Isolation (Partial)

	Rat Cardiac Microvascular Endothelial Cells (CD31)	Rat Cardiac Fibroblasts (Vimentin)	Mouse Cardiomyocytes (α-SCA)	Mouse Cardiac Fibroblasts (Vimentin)
Microscopic View of Cell Morphology
Immunofluorescence Identification

Primary Cardiac-Derived cells 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 Cardiac-Derived Cell Isolation and Culture

1.Do cardiomyocyte and cardiac fibroblast isolations require neonatal mice and rats (1-2 days old), or can older mice be used?

When choosing the age of mice and rats, neonatal mice and rats within the first 1-2 days after birth are generally recommended. This is because cardiomyocyte differentiation progresses rapidly with age. Neonatal cardiomyocytes still retain some proliferative capacity, whereas adult cardiomyocytes are terminally differentiated, have poor adherence, low viability, and completely lack proliferative ability. Fibroblast proliferation capacity also varies with age. For these reasons, neonatal mice and rats are preferred, as they yield cells with higher survival and adherence rates.

Isolating cardiomyocytes from adult mice and rats is possible but requires the Langendorff retrograde perfusion technique. This approach is technically demanding and time-consuming, and it typically yields fewer cells with lower viability, making it unsuitable for large-scale experiments.

2.How can contamination between cardiomyocytes and cardiac fibroblasts be minimized?

It is true that these two cell types are often mixed during isolation. To separate them, researchers commonly use differential adhesion and differential digestion techniques. Fibroblasts adhere more quickly than cardiomyocytes, allowing preliminary purification based on adhesion time. The recommended timing provided in the Pricella® product manual reflects optimized conditions, but users can adjust it as needed for their specific experiments to improve cell purity.

3. After culturing cardiac microvascular endothelial cells, impurities adhering to the cell surface. Is this normal?

Yes, this is normal. When removing impurities, avoid rinsing the cells vigorously with culture medium or PBS, as this can easily cause detachment. Instead, remove the culture supernatant, add PBS, and gently tap the bottom of the culture dish to allow the impurities to fall off naturally. Then discard the PBS and replace it with fresh culture medium.

4.Can cardiac-derived cells be subcultured, and which digestive enzymes should be used?

Cardiac fibroblasts: These cells have relatively strong proliferative capacity and can be subcultured using trypsin. They can generally be passaged several three times.

Cardiac microvascular endothelial cells: Their proliferation is weaker than that of fibroblasts. The Accutase Cell Detachment Solution is recommended for subculture. Rat-derived cells can typically be passaged about three times, while mouse-derived cells are best limited to 2-3 passages. Extensive subculture is not recommended.

Cardiomyocytes: These are terminally differentiated cells with no proliferative ability. If enzymatic digestion is required during culture, trypsin can be used, though this may negatively affect cell viability.

5.Are there morphological differences between cardiac-derived cells from rats and mice?

Cardiac fibroblasts: Morphological differences between rat and mouse fibroblasts are minimal.

Cardiomyocytes: Rat cardiomyocytes beat more strongly and frequently than mouse cardiomyocytes.

Cardiac microvascular endothelial cells: Mouse endothelial cells often display a more irregular morphology and a less pronounced “cobblestone” arrangement compared with those of rat endothelial cells. In addition, they can typically be subcultured fewer times.