Long-term storage of PRP: progress, stability and the future of regenerative medicine

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  • PRP storage, Cryopreservation, Regenerative Medicine, Growth Factors
Long-term storage of PRP: progress, stability and the future of regenerative medicine

Shelf life of PRP: processing time, storage conditions and biological changes

The composition of PRP makes it a sensitive product. Platelets and growth factors such as PDGF (platelet-derived growth factor), VEGF (vascular endothelial growth factor) and TGF-β (transforming growth factor-beta) are bioactive molecules that can degrade or become inactive if stored improperly. In addition, the storage of PRP can lead to an uncontrolled release of growth factors, which reduces the effectiveness of the application.

1. How quickly must PRP be processed after centrifugation?

After obtaining PRP by centrifugation, time is a critical factor. The platelets, which are the main source of growth factors, only remain active and intact for a limited period of time. If PRP is not processed or applied in time, the platelets lose their function and the growth factors begin to degrade.

1.1 Processing time

  • Unrefrigerated (room temperature, 20-25 °C):
    • PRP should ideally be used within 30 minutes to a maximum of 2 hours after centrifugation to ensure full bioactive efficacy.
    • After 2 hours, platelet activation begins spontaneously, leading to an uncontrolled release of growth factors. These growth factors are only stable for a limited period of time before they are degraded.

  • Refrigerated (4 °C):
    • When stored at 4 °C, PRP can remain stable for up to 4-6 hours. Cooling slows down platelet metabolism and reduces spontaneous activation.
    • However, prolonged storage at 4 °C leads to a decrease in growth factors and impaired platelet integrity.

2. Biological processes in PRP after centrifugation

After the production of PRP, various biological processes begin that influence its stability and effectiveness. These processes affect both the platelets and the dissolved growth factors and proteins.

2.1 Activation of the thrombocytes

  • After centrifugation, the platelets are in a dormant state.
  • Without appropriate stimulation (e.g. by calcium chloride or thrombin), the platelets can activate spontaneously within a few hours.

  • Consequence of activation:
    • Release of growth factors such as PDGF (platelet-derived growth factor), VEGF (vascular endothelial growth factor) and TGF-β (transforming growth factor-beta) in an uncontrolled manner.
    • After release, these growth factors begin to degrade rapidly, which impairs the therapeutic efficacy of PRP.

2.2 Degradation of growth factors

  • The growth factors that are released after platelet activation are sensitive proteins that are degraded by enzymes and oxidative stress.
  • At room temperature, these factors can lose a significant amount of their concentration within 4-6 hours.

2.3 Change in cell components

The platelet membranes begin to destabilize during prolonged storage, especially at temperatures above 4 °C. This leads to:

  • Loss of cell integrity.
  • Release of intracellular enzymes that further impair the stability of the PRP components.

2.4 Oxidative and enzymatic processes

  • Oxidative stress (caused by oxygen and free radicals) can damage proteins and lipids in the PRP.
  • Enzymes such as proteases and peroxidases, which are present in the plasma, contribute to the degradation of growth factors and proteins.

3. Shelf life of PRP under different storage conditions

The storage of PRP is crucial in order to maintain its bioactive efficacy. Here are the different storage conditions and their influence on shelf life:

3.1 Unrefrigerated (room temperature, 20-25 °C):

  • Shelf life: 30 minutes to 2 hours.
  • At room temperature, spontaneous platelet activation starts quickly, leading to a loss of bioactive substances.
  • After 2 hours, most of the growth factors are released and begin to degrade.

3.2 Refrigerated (4 °C):

  • Shelf life: 4-6 hours.
  • Cooling slows down the metabolism of the platelets and reduces activation.
  • The release and degradation of growth factors are slowed down, but PRP does not remain stable for more than 6 hours at 4 °C.

3.3 Frozen (-20 °C):

  • Shelf life: Up to 3 months.
  • Freezing at -20 °C maintains the concentration of growth factors for several weeks to months. However, platelets can be damaged by the formation of ice crystals, which impairs their integrity.

3.4 Deep-frozen (-80 °C):

  • Shelf life: 6-12 months.
  • At -80 °C, most growth factors and proteins remain stable. However, platelets lose their cell integrity and thus the ability to actively release growth factors.
  • PRP stored at -80 °C is particularly suitable for applications in which the growth factors in the plasma itself are used (e.g. in the treatment of tissue injuries).

3.5 Cryopreservation (-196 °C, liquid nitrogen):

  • Shelf life: 12 months or longer.
  • Cryopreservation at -196 °C is the best method to ensure the long-term stability of PRP. It preserves the bioactive functionality of the growth factors and reduces damage to the platelets through the use of cryoprotectants such as DMSO.

4. Recommendations for practice

The storage of PRP is crucial in order to maintain its bioactive efficacy. Here are the different storage conditions and their influence on shelf life:

4.1 Processing and application of freshly prepared PRP

  • Prepare PRP as soon as possible after centrifugation and use it within 2 hours for optimal results.
  • If immediate use is not possible, store PRP in the refrigerator at 4 °C for no longer than 6 hours.

4.2 If storage is required

  • Use appropriate storage methods (e.g. -20 °C or lower) if PRP is to be preserved for later use.
  • If PRP is frozen, ensure that cryoprotectants are used to minimize cell damage from ice crystals.

4.3 Thawing stored PRP

  • PRP should be thawed gently, e.g. at room temperature or in a water bath at a maximum of 37 °C.
  • Avoid rapid thawing at high temperatures, as this can damage the proteins and platelets.

5. Conclusion

The shelf life of PRP is highly dependent on the storage conditions and the time that elapses between production and application. Ideally, freshly prepared PRP should be used within 2 hours to ensure full bioactive efficacy. For longer storage, refrigerated or frozen conditions (4 °C to -80 °C) offer a viable solution. Frozen PRP at -80 °C or cryopreserved at -196 °C can remain stable for months, but is particularly suitable for planned follow-up treatments. Careful handling and documentation are crucial to ensure the quality of the PRP and thus the success of the therapy.

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