Pricella® Primary Bone Marrow-Derived Cell Isolation Kits: Precise Isolation of Multilineage Cells for Advanced Bone Marrow Research

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

Ⅰ. Primary Bone Marrow-Derived Cell Types and Their Research

Bone marrow is an important hematopoietic and immune-regulating organ in the body. It contains various types of precursor cells and immune cells, and is involved in multiple physiological processes, such as tissue repair, immune modulation, and angiogenesis. In research, the commonly used bone marrow-derived cells mainly include the following categories:

Bone Marrow Monocytes: They refer to the immature monocyte population in the bone marrow, accounting for approximately 0.04% of total bone marrow cells. They give rise to multiple immune-related cell types (such as osteoclasts, macrophages) and play a key role in regulating inflammation and maintaining the immune microenvironment. Therefore, they are widely used in disease modeling and the study of therapeutic strategies.

Bone Marrow Mononuclear Cells: They refer to a cell population with a single nucleus, including lymphocytes, monocytes, hematopoietic stem/progenitor cells, and other progenitor cells. Since they contain a variety of functional cells, they possess multiple capabilities, such as immune regulation, promotion of angiogenesis, and tissue repair. They are an important cellular source for studying the regeneration mechanisms after tissue injury.

Bone Marrow-Derived Macrophages: They differentiate from bone marrow monocytes under specific induction conditions, and possess strong phagocytic ability and high plasticity. They can present different polarization states (such as pro-inflammatory M1 type and anti-inflammatory M2 type) under different stimuli. They are often used to model dynamic macrophage responses under pathological conditions and are important tools for studying chronic inflammation, autoimmune diseases, and the tumor immune microenvironment.

Bone Marrow-Derived Endothelial Progenitor Cells: They are a group of early endothelial-like cells with migratory ability and the potential for further proliferation and differentiation. Usually, they lack the typical phenotypic markers of mature endothelial cells. Their main function is to participate in postnatal angiogenesis in ischemic tissues and in vascular repair after injury. They are ideal cellular models for exploring the mechanisms of endogenous vascular regeneration.

Ⅱ. Pricella^® Primary Bone Marrow-Derived Cell Isolation Kits

1.  Product Catalog 

2. Kit Components and Brief Description of Their Functions

Washing Solution: Used for rinsing femoral and tibial tissues, effectively removing impurities and maintaining cell activity, and serves to inhibit bacterial growth.

Specialized Lysis Solution (Bone Marrow-Derived Macrophages/Endothelial Progenitor Cells): Used to remove red blood cells to improve target cell purity and reduce background interference.

Specialized Isolation Solution (Bone Marrow Monocytes/Mononuclear Cells): Based on differences in cell density, this solution enriches target cells, thereby enabling separation and purification.

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.

Primary bone marrow-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.

3.Cell Images from Kit Isolation (Partial)

	Rat Bone Marrow-Derived Endothelial Progenitor Cells (CD34)	Rat Bone Marrow-Derived Macrophages (CD68)	Rat Bone Marrow Monocytes (CD68)	Mouse Bone Marrow-Derived Macrophages (CD68)
Microscopic View of Cell Morphology
Immunofluorescence Identification

Primary bone marrow-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 Bone Marrow-Derived Cell Isolation and Culture

1. Insufficient Cell Purity 

During sample collection, the muscle and tendon tissues were not fully stripped, and their fragments were exposed in the bone marrow flushing fluid, reducing cell purity. It is recommended to completely remove the muscle and tendon tissues of the experimental mice during the operation.

If the bone marrow fluid is exposed to the muscle tissue after the bone is cut, the corresponding bone tissue should be discarded to avoid cell contamination. If the cell number is insufficient, increase the number of animals as needed.

2. Low Yield of Bone Marrow-Derived Monocytes and Mononuclear Cells 

Incomplete flushing of the bone marrow can reduce cell yield. Flush thoroughly until the bone cavity appears white.

If a large amount of tissue is discarded due to improper bone cutting or breaking, the number of animals for extraction can be appropriately increased.

During the separation process, if the stratification is not obvious, it may affect the effective separation of cells. It is suggested that when slowly dropping the cell suspension onto the separation medium, avoid disturbing the liquid surface, which may cause unclear stratification and thus reduce separation efficiency.

3. Slow Adhesion of Bone Marrow-Derived Endothelial Progenitor Cells 

Different from other primary cells, bone marrow-derived endothelial progenitor cells have a relatively long culture period, requiring at least 10-14 d. During the culture period, it is recommended to centrifuge and replace the medium every 2-3 d, and then transfer the cells back to the original dish for continued culture.

The adhesion efficiency is greatly affected by the age of the animals. It is recommended to strictly follow the mouse age specified in the instruction manual for material collection.

4. Difficulty in Digesting Adherent Bone Marrow-Derived Macrophages and Monocytes 

Macrophages and monocytes adhere firmly and are more difficult to digest than other primary cells. It is recommended to directly seed the cells into culture vessels for experiments after isolation and to avoid repeated digestion whenever possible.

If digestion is necessary, refer to the digestion protocol recommended in the instruction manual and adopt the “multiple short-duration” digestion method to prevent the cells from being exposed to the digestive fluid for too long.