Key Takeaways & Executive Findings
- •• Optimized whole blood separation process enables production of therapeutic-grade platelet lysate with high concentrations of growth factors. • Platelet lysate prepared by freeze-thawing is free from bacterial and mycoplasma contamination, ensuring safety for potential clinical use. • Low concentration (1%) platelet lysate significantly enhances cardiomyocyte proliferation and protects against hypoxic injury by upregulating antioxidant enzymes (SOD and GSH-Px). • Platelet lysate shows promise as a cost-effective and efficient supplement for cardiac tissue engineering and regenerative medicine.
Abstract
BACKGROUND: Platelets are important blood resources, yet in routine blood bank processes they are often filtered out along with white blood cells as medical waste. Optimizing whole blood separation processes to prepare platelet lysate products and exploring their applications in tissue engineering and regenerative medicine is of great value. OBJECTIVE: To optimize whole blood separation to prepare therapeutic-grade platelet lysate and to investigate the protective effect of platelet lysate on hypoxic injury of cardiomyocytes. METHODS: Platelets were isolated from 21 qualified whole blood units under closed blood bag and tubing conditions, and 21 platelet lysates were prepared by freeze-thawing. The mass concentration ranges of platelet-derived growth factor AA, platelet-derived growth factor BB, platelet-derived growth factor AB, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor 1, fibroblast growth factor, and transforming growth factor beta 1 in platelet lysates were measured using enzyme-linked immunosorbent assay kits. Bacterial contamination was assessed by colony culture method and mycoplasma contamination by PCR detection kit. A cardiomyocyte hypoxia model was established to evaluate the protective effect of platelet lysate on hypoxic injury. RESULTS AND CONCLUSION: (1) The mass concentration ranges of major growth factors and cytokines in platelet lysates were: platelet-derived growth factor AA 12.86-24.17 μg/L, platelet-derived growth factor BB 0.25-0.32 μg/L, platelet-derived growth factor AB 85.09-114.91 μg/L, vascular endothelial growth factor 10.57-58.37 μg/L, epidermal growth factor 0.43-0.69 μg/L, insulin-like growth factor 1 106-204.9 μg/L, fibroblast growth factor 0.03-0.06 μg/L, and transforming growth factor beta 1 124.17-192.38 μg/L. (2) Colony culture and mycoplasma detection results were negative. (3) Low volume fraction (1%) platelet lysate yielded the highest proliferation efficiency of cardiomyocytes; low volume fraction (1%) platelet lysate stimulated cardiomyocytes to produce high levels of superoxide dismutase and glutathione peroxidase to protect cardiomyocytes. This study established a method for preparing therapeutic-grade platelet lysate by optimizing the whole blood separation process, which can improve the utilization rate of blood resources. Platelet lysate has high levels of major growth factors and can significantly promote the repair of hypoxic injured cardiomyocytes.
1. Introduction
Concentrated platelet products are widely used in transfusion medicine to prevent or treat bleeding disorders caused by thrombocytopenia or platelet dysfunction. In recent years, regenerative medicine and tissue engineering have focused on the development of platelet derivatives, such as platelet-rich plasma, platelet-rich fibrin, and platelet lysate, due to their rich content of growth factors and bioactive molecules. Platelet lysate, as a third-generation platelet derivative, contains higher levels of growth factors and bioactive components and can be stored at 4-10 °C for 14 days or frozen for longer periods, overcoming the limitations of platelet-rich plasma, which is temperature-sensitive and has a short shelf life. Platelet lysate is easier to standardize in preparation, making it a ready-to-use alternative medical product with broad clinical application prospects. However, in routine blood bank processes, platelets are often filtered out along with white blood cells as medical waste, leading to waste of platelet resources. In the current context of blood resource shortage, it is of great significance to fully utilize platelet resources from whole blood to prepare platelet derivatives.
During myocardial infarction, insufficient blood supply to the heart causes oxidative stress in cardiomyocytes, leading to metabolic abnormalities, structural damage, and functional impairment due to inadequate oxygen supply or utilization, resulting in hypoxic injury and subsequent apoptosis. To date, no studies have reported the application of platelet lysate in cardiomyocyte culture. Therefore, research on platelet lysate in the field of cardiomyocyte proliferation and protection is of great value and exploratory significance. Platelet lysate exerts protective effects on hypoxic injured cardiomyocytes mainly through abundant growth factors, cytokines, and bioactive substances. Additionally, platelet lysate can effectively alleviate hypoxic injury and promote tissue repair through multi-target synergistic actions, including promoting angiogenesis, anti-apoptosis, anti-inflammation, anti-oxidation, and metabolic regulation. Thus, the natural combination of components in platelet lysate offers advantages over single factors in treatment. Systematic investigation of the protective effects and mechanisms of platelet lysate on hypoxic injured cardiomyocytes will provide new strategies for the treatment of myocardial infarction and other ischemic heart diseases.
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Jiang Lihong, Lin Fuwen, Chen Ying, Huang Yuchen, Peng Shaojing, Chen Jierun, Su Changshan, Zhong Zhoulin (2026). Protective effect of optimization of the whole blood separation process to prepare therapeutic-grade platelet lysate on cardiomyocytes from hypoxic injury. Chinese Journal of Tissue Engineering Research. https://doi.org/10.12307/2026.21348
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Frequently Asked Questions
What is platelet lysate and how is it prepared?
Platelet lysate is a liquid prepared by lysing platelets and collecting the released intracellular components. It is rich in growth factors such as platelet-derived growth factor, vascular endothelial growth factor, epidermal growth factor, and transforming growth factor beta. In this study, platelet lysate was prepared by freeze-thawing platelets isolated from whole blood under closed conditions.
What are the key growth factors in platelet lysate and their concentrations?
The platelet lysate contains high concentrations of growth factors including platelet-derived growth factor AA (12.86-24.17 μg/L), platelet-derived growth factor BB (0.25-0.32 μg/L), platelet-derived growth factor AB (85.09-114.91 μg/L), vascular endothelial growth factor (10.57-58.37 μg/L), epidermal growth factor (0.43-0.69 μg/L), insulin-like growth factor 1 (106-204.9 μg/L), fibroblast growth factor (0.03-0.06 μg/L), and transforming growth factor beta 1 (124.17-192.38 μg/L).
How does platelet lysate protect cardiomyocytes from hypoxic injury?
Platelet lysate protects cardiomyocytes from hypoxic injury by promoting cell proliferation and upregulating antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. These enzymes reduce oxidative stress and apoptosis, thereby enhancing cell survival and repair.
What is the optimal concentration of platelet lysate for cardiomyocyte culture?
The study found that a low volume fraction (1%) of platelet lysate resulted in the highest proliferation efficiency of cardiomyocytes and stimulated high levels of protective enzymes, indicating that 1% is the optimal concentration for cardiomyocyte culture.
What are the potential applications of platelet lysate in regenerative medicine?
Platelet lysate has potential applications in tissue engineering and regenerative medicine, particularly for cardiac repair after myocardial infarction. Its high growth factor content and ability to promote cell proliferation and protect against injury make it a promising supplement for cell culture and therapeutic applications.
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