The issue of clot formation in cellular studies with platelet-rich plasma and ways to its solutions

Abstract

Summary. Introduction. Clot formation in a culture medium with added platelet-rich plasma (PRP) significantly complicates the study of the functional cell properties. The need for an effective solution to this issue is attributed to its impact on the assessment of cell viability and metabolism, which jeopardizes the validity of the experimental results. Aim: to study the effect of various agents on clot formation in PRP and to evaluate their impact on the viability and metabolism of human skin fibroblasts (HSF). Materials and Methods. Blood samples from five volunteer donors (median age 20 [19; 21] years) were used for the study. The standard procedure of double centrifugation was utilized for preparing PRP. Two steps of PRP activation were performed: thawing followed by the addition of an agent and calcium chloride. Before activation and addition to the culture medium, various chemical substances were added to each sample to prevent clot formation: fibrin monomer polymerization inhibitor (FMPI), heparin, and dextran sulfate in different concentrations. The viability of the hTERT-HDFa cells (immortalized human skin fibroblast cells) was determined by the MTT assay (colorimetric assay for assessing cell metabolic activity) and the Alamar Blue assay. Results. The data obtained showed that heparin at a concentration of 25 IU/ml and FMPI at a concentration of 1 mg/ml are the most effective in preventing clot formation and improving cell viability and metabolism compared to the control group. Conclusion. The most effective agents for preventing clot formation are heparin and FMPI. The obtained results require additional confirmation and further studies to identify the mechanisms of improving cell viability when using them.

For citation: Vlasova T.I., Momot A.P, Brodovskaya E.P., Belozerskaya G.G., Madonov K.S., Bezborodova A.P., Abelova A.P., Polozova A.I. The issue of clot formation in cellular studies with platelet-rich plasma and ways to its solution. Tromboz, gemostaz i reologiya. 2026;(2):55–62. (In Russ.).

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