A Safe and Convenient Method to Isolate Bone Marrow Mononuclear Cells in Clinical Practice

Area of Science:

• Regenerative Medicine
• Cell Therapy
• Hematology

Background:

• Bone marrow mononuclear cells (BMMNCs) are crucial for regenerative medicine.
• Current isolation methods like density gradient centrifugation (DGC) are complex and operator-dependent.

Purpose of the Study:

• To develop a simple, safe, and efficient method for isolating BMMNCs.
• To optimize hypotonic sodium chloride (NaCl) concentration for BMMNC isolation.

Main Methods:

• Investigated optimal hypotonic NaCl concentration for BMMNC viability and lysis efficiency in minipigs.
• Compared 0.3% NaCl lysis with ammonium chloride (NH4Cl) lysis and DGC.
• Assessed nucleated cell yield, residual red blood cells (RBCs), BMMNC viability, apoptosis, and colony-forming ability.

Main Results:

• 0.3% NaCl lysis demonstrated optimal viability and lysis efficiency.
• The 0.3% NaCl method yielded more cells and had fewer residual RBCs than DGC.
• Bone marrow mesenchymal stem cells (BMSCs) isolated via NaCl lysis showed comparable biological characteristics to other methods, except for proliferation which was best in the NaCl group.

Conclusions:

• 0.3% NaCl is the optimal concentration for hypotonic lysis of BMMNCs.
• 0.3% NaCl lysis is a safe, appropriate, and cost-effective alternative to NH4Cl lysis and DGC for BMMNC isolation.