Home › Cell Biology › A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells
Cell Biology JoVE (Open Access) Citable · DOI

A Simple and Efficient Method to Isolate Macrophages from Mixed Primary Cultures of Adult Liver Cells

DOI: 10.3791/2757-v
What you'll learn
  • ✓ Perform in situ liver perfusion to isolate primary hepatocytes
  • ✓ Culture mixed liver cell populations and selectively expand macrophages
  • ✓ Purify macrophages using differential attachment and shaking techniques
  • ✓ Obtain high-purity liver macrophages without specialized equipment
Protocol

Biopharma Insights A novel method to obtain macrophages from primary culture of rat liver cells is described. This method utilizes the proliferation of macrophages in the culture, followed by shaking of culture flasks and purification by selective attachment to plastic dishes. This technique efficiently provides liver macrophages without complex equipment and skills.

Difficulty
intermediate
Total time
~3–4 hours per rat liver (perfusion through final purification)
Model organism
Rat (adult, species unspecified)
Biosafety
BSL-1

Steps

1
Perfuse liver tissue with buffer solution

Perform in situ perfusion of rat liver using appropriate buffer to remove blood and prepare tissue for cell isolation. This step is critical for obtaining viable primary cells.

▶ 02:14
2
Prepare hepatocyte-rich primary cell fraction

Dissociate perfused liver tissue and prepare a mixed primary culture enriched for hepatocytes. This heterogeneous culture serves as the starting material for selective macrophage isolation.

▶ 03:33
3
Selectively isolate macrophages by differential attachment

Culture the mixed cell population and exploit macrophage proliferation and selective plastic adhesion. Shake culture flasks to dislodge non-adherent cells, then transfer supernatant to fresh plates to enrich adherent macrophages.

▶ 04:36
4
Verify macrophage purity and viability

Assess the final isolated macrophage population for purity and morphological characteristics using microscopy or immunological markers. Results demonstrate highly purified rat liver macrophages suitable for downstream applications.

▶ 06:46
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