An enzymatic method to rescue mesenchymal stem cells from clotted bone marrow samples

Area of Science:

• Regenerative Medicine
• Stem Cell Biology
• Hematology

Background:

• Mesenchymal stem cells (MSCs) are crucial for tissue engineering and autologous therapies.
• Bone marrow aspirates can form clots upon collection, leading to significant loss of viable MSCs.
• Clotted samples reduce stem cell yield by over 50%, necessitating additional procedures.

Purpose of the Study:

• To develop a cost-effective protocol for recovering MSCs from clotted bone marrow samples.
• To evaluate the efficacy of enzymatic clot digestion using urokinase.
• To ensure the quality and functionality of recovered MSCs are maintained.

Main Methods:

• A protocol was developed utilizing clinical-grade urokinase for enzymatic digestion of bone marrow clots.
• Canine bone marrow samples were treated with urokinase to assess clot degradation and MSC recovery.
• Metabolic activity and differentiation potential of urokinase-treated MSCs were analyzed.

Main Results:

• Urokinase treatment effectively degraded bone marrow clots, recovering a comparable number of MSCs to unclotted samples.
• 74% of canine bone marrow samples contained clots, with clotted samples yielding less than half the expected MSCs.
• MSCs recovered after urokinase treatment retained metabolic activity and differentiation capacity.

Conclusions:

• Enzymatic digestion with urokinase salvages clotted bone marrow samples, maximizing MSC yield.
• This protocol offers a rapid, inexpensive alternative to bone marrow resampling, reducing costs and preserving cell quality.
• The method enables the use of previously discarded clotted samples for research and therapeutic purposes.