# Optimized Centrifugation and Activation Protocol for the Preparation of Plasma Rich in Growth Factors in Pigs

## TL;DR

Researchers developed a reliable method to prepare growth factor-rich plasma in pigs, which could help improve cartilage repair studies.

## Contribution

The study introduces a standardized protocol for porcine plasma rich in growth factors (PRGF) aligned with human clinical practices.

## Key findings

- A 400× g centrifugation and 13.3 mM CaCl2 activation protocol yielded optimal platelet recovery and fibrin network formation.
- Porcine PRGF showed similar growth factor profiles to human PRGF when using the same preparation protocol.
- The protocol supports the pig as a relevant model for preclinical evaluation of regenerative cartilage therapies.

## Abstract

Background: Cartilage defects remain a clinical challenge due to the limited intrinsic repair capacity of hyaline cartilage, driving increasing interest in blood-derived products, including platelet-rich plasma (PRP). Variability in PRP preparation and activation protocols limits reproducibility and clinical translation, particularly in large animal models where species-specific differences are an additional cue. This study aimed to standardize and optimize in pigs a protocol for plasma rich in growth factors (PRGF), a leukocyte-poor PRP, aligned with current human clinical practice. Methods: Whole blood from six female pigs was processed via three centrifugation protocols and activated with varying CaCl2 concentrations to evaluate gelation and morphology. PRGF was characterized through hematological analysis, ELISA-based quantification of soluble factors, and structural imaging of fibrin gel via histology and scanning electron microscopy. Data were further analyzed using protein–protein interaction networks, hierarchical clustering, and comparative human PRGF proteomic profiles. Results: Protocol with 400× g centrifugation followed by 13.3 mM CaCl2 activation achieved the most favorable performance, yielding the highest platelet recovery, effective leukocyte clearance, and consistent formation of a well-organized fibrin network. Porcine activated PRGF showed substantial overlap in detected factors and concentration ranges with human activated PRGF prepared with the same protocol. Conclusions: These findings establish a robust, clinically aligned porcine PRGF protocol and support the pig as a relevant translational model for PRP-based regenerative strategies, providing a reliable platform for preclinical evaluation of cartilage therapies.