Protective effect of optimization of the whole blood separation process to prepare therapeutic-grade platelet lysate on cardiomyocytes from hypoxic injury

doi:10.12307/2026.223

Abstract

Abstract: BACKGROUND: Platelets, crucial blood components, are commonly discarded along with white blood cells in traditional blood processing and become medical waste. Optimizing whole blood separation processes to prepare platelet lysate products and exploring their applications in tissue engineering and regenerative medicine are of great value.
OBJECTIVE: To optimize blood separation for therapeutic-grade platelet lysate production and explore the protective impact of platelet lysate on myocardial hypoxia injury.
METHODS: Twenty-one platelets were isolated from whole blood using a closed blood bag and tubing system, followed by the preparation of twenty-one platelet lysates through the freeze-thaw method. The concentrations of platelet-derived growth factors (platelet-derived growth factor AA, platelet-derived growth factor BB, and platelet-derived growth factor AB), vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor, fibroblast growth factor, and transforming growth factor-β1 in the platelet lysate were quantified using an enzyme-linked immunosorbent assay kit. Detection ranges for these factors were determined. Bacterial and mycoplasma contamination were observed using the colony culture method and a Mycoplasma PCR detection kit, respectively. A hypoxic cardiomyocyte model was established to evaluate the protective effects of platelet lysate on hypoxia-stressed cardiomyocytes.
RESULTS AND CONCLUSION: (1) The concentration ranges of major growth factors and cytokines in platelet lysates were as follows: 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, transforming growth factor β1: 124.17-192.38 μg/L. (2) Colony culture and mycoplasma detection yielded negative results. (3) The efficiency of cardiomyocyte proliferation was highest with a low concentration (1%) of platelet lysate in culture. A 1% concentration of platelet lysate effectively stimulated cardiomyocytes to produce high levels of superoxide dismutase and glutathione peroxidase, providing protective effects for cardiomyocytes. This study established a method for preparing therapeutic-grade platelet lysate by optimizing the whole blood separation process, thereby improving the utilization of blood resources. Platelet lysate, rich in essential growth factors, significantly promotes the repair of hypoxic injuries in cardiomyocytes.

Key words: platelet lysate, platelet-rich plasma, growth factor, cell culture, myocardial cell, superoxide dismutase, glutathion peroxidase

CLC Number:

Jiang Lihong, Lin Fuwen, Chen Ying, Huang Yuchen, Peng Shaojing, Chen Jierun, Su Changshan, Zhong Zhoulin. Protective effect of optimization of the whole blood separation process to prepare therapeutic-grade platelet lysate on cardiomyocytes from hypoxic injury[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(19): 4983-4989.

Figures/Tables 7

2.5 低体积分数血小板裂解液可促进缺氧损伤人心肌细胞增殖各组间细胞总数存在显著差异(P < 0.001)。对于血小板裂解液，体积分数1%组细胞总数显著高于体积分数5%组和体积分数10%组(均P < 0.001)，且体积分数5%组细胞总数显著高于体积分数10%组(P < 0.001)。对于富血小板血浆，体积分数5%组显著高于体积分数1%组和体积分数10%组(均P < 0.001)。对于新鲜血浆，体积分数5%组显著高于体积分数1%组和体积分数10%组(均P=0.024)。当体积分数控制在1%时，血小板裂解液组细胞总数显著高于富血小板血浆组和新鲜血浆组(均P < 0.001)。当体积分数控制在5%时，新鲜血浆组细胞总数显著低于血小板裂解液组(P=0.003)和富血小板血浆组(P=0.006)。当体积分数控制在10%时，富血小板血浆组细胞总数显著高于血小板裂解液组(P < 0.001)和新鲜血浆组(P=0.008)，而新鲜血浆组显著高于血小板裂解液组(P < 0.001)。各组间细胞存活率存在显著差异(P < 0.001)。对于血小板裂解液，体积分数1%组细胞存活率显著高于体积分数5%组和10%组(均P < 0.001)，且体积分数5%组细胞存活率显著高于体积分数10%组(P < 0.001)。对于富血小板血浆，体积分数10%组细胞存活率显著高于体积分数1%组和体积分数5%组(均P < 0.001)，且体积分数5%组细胞存活率显著高于体积分数1%组(P < 0.001)。对于新鲜血浆，体积分数10%组细胞存活率显著高于体积分数1%组和体积分数5%组(均P < 0.001)，且体积分数5%组细胞存活率显著高于体积分数1%组(P < 0.001)。当体积分数控制在1%时，血小板裂解液组细胞存活率显著高于富血小板血浆组和新鲜血浆组(均P < 0.001)，而富血小板血浆组细胞存活率显著低于新鲜血浆组(P < 0.001)。当体积分数控制在5%时，血小板裂解液组细胞存活率显著高于富血小板血浆组和新鲜血浆组(均P < 0.001)，而富血小板血浆组细胞存活率显著低于新鲜血浆组(P < 0.001)。当体积分数控制在10%时，富血小板血浆组细胞存活率显著高于血小板裂解液组和新鲜血浆组(均P < 0.001)，而血小板裂解液组细胞存活率显著低于新鲜血浆组(P < 0.001)。因此，根据细胞总数和细胞存活率的比较得出：体积分数1%血小板裂解液修复损伤心肌细胞的能力以及促进细胞增殖的效率均显著优于富血小板血浆和新鲜血浆。低体积分数血小板裂解液组的细胞总数和细胞存活数均高于富血小板血浆组和新鲜血浆组，这进一步证实了低体积分数血小板裂解液在心肌细胞损伤修复和增殖中的优势，见表2。 "

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