Mouse Bone Marrow Cell Isolation Protocol

This mouse bone marrow isolation protocol outlines the detailed steps for isolating mouse bone marrow cells. Unlike whole blood, mouse bone marrow yields a diverse set of cells, including hematopoietic stem cells responsible for producing leukocytes, erythrocytes, and thrombocytes, as well as stromal cells such as endothelial cells, fibroblasts, macrophages, osteoblasts, osteoclasts, and adipocytes. This comprehensive method helps ensure the successful isolation of a diverse cell population for various research applications. Evaluation methods such as flow cytometry, ELISA, and cell culture assays can be used to analyze the isolated mouse bone marrow cells, helping provide valuable insights into hematopoiesis and stromal cell functions.

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• Cell culture dishes (e.g., Thermo Scientific Nunc EasYDish Dishes, 100 mm, Cat. No. 150466)
• Lab wipes (e.g., Fisher Scientific Kimberly-Clark Professional Kimtech Science Kimwipes Delicate Task Wipers, 1-PlyFisher, Fisher Scientific Cat. No. 06-666)
• Ethanol (e.g., Absolute Ethanol, 200 Proof, Molecular Biology Grade, Cat. No. T038181000CS)
• RPMI Complete Media
  • RPMI 1640 medium (e.g., Gibco BenchStable RPMI 1640, Cat. No. A4192301)
  • 2 mM L-glutamine (e.g., Gibco L-Glutamine, 200 mM, Cat. No. 25030149)
  • Fetal bovine serum (e.g., Gibco Fetal Bovine Serum, Cat. No. 26140079, or Fetal Bovine Serum One Shot format, Cat No. A3160401)
  • Optional: Penicillin-streptomycin (e.g., Gibco Penicillin-Streptomycin (5,000 U/mL), Cat. No. 15070063, or Gibco Penicillin-Streptomycin (10,000 U/mL), Cat. No. 15140122)
• Scalpel and blades
• 5–10 mL syringe (e.g., Fisherbrand Sterile Syringes for Single Use, 10 mL, Cat. No. 14-955-459)
• 25-gauge needle (e.g., Fisher Scientific BD General Use and PrecisionGlide Hypodermic Needles, Cat. No. 14-826-49)
• 70 μm cell strainers (e.g., Fisherbrand Sterile Cell Strainers, Cat. No. 22-363-548)
• 50 mL conical tubes (e.g., Thermo Scientific Nunc 50 mL Conical Sterile Polypropylene Centrifuge Tubes, Cat. No. 339652)
• Red blood cell (RBC) lysis buffer (e.g., Invitrogen eBioscience 1X RBC Lysis Buffer, Cat No. 00-4333-57 or Invitrogen eBioscience 10X RBC Lysis Buffer (Multi-species), Cat. No. 00-4300-54)
• Cell counter (e.g., Invitrogen Countess 3 Automated Cell Counter, Cat. No. AMQAX2000)
• Phosphate-buffered saline (PBS) (e.g., Gibco PBS (10X), pH 7.4, Cat. No. 70011069)

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