In modern biomedicine, there is an increasingly urgent need for high-resolution data on the fates of individual cells during ontogenesis from the zygote to the formation of mature tissues particularly for complex organs such as the heart, brain, and spinal cord. A detailed understanding of these processes will enable the development of novel approaches in tissue and cell engineering, and open new avenues for correcting previously incurable diseases, including neurodegenerative, cognitive, and mental. Among the most informative tools for reconstructing cell lineage are genetic barcoding technologies, which are the focus of this review. In this literature survey, we trace the evolution of genetic barcoding from early fluorescent systems (Brainbow, Confetti) to modern DNA-barcoding platforms based on programmable nucleases Cas9 and Cas12a (GESTALT, MEMOIR, CARLIN, DuTracer, and others). We analyze and compare their key properties, capabilities, and limitations, as well as their potential applications in developmental biology and regenerative biomedicine. Further advancements in genetic barcoding technologies hold promise for generating highly informative disease models, validating pharmacological targets, and developing cell-based therapeutics for regenerative medicine. We anticipate that this review will be of broad interest to a diverse audience including developers of cell and genetic technologies, researchers in regenerative biomedicine and developmental biology, omics specialists, and investigators studying age-related, neurodegenerative, and oncohematological diseases by providing a comprehensive overview of a foundational technology for reconstructing cell lineage to address both fundamental and applied challenges across a wide spectrum of biological and medical sciences.

This article presents a risk-based approach to the development of a quality control strategy for biological medicinal products, with a particular emphasis on the role of the analytical control strategy as a central element of an integrated quality assurance system. It is demonstrated that the inherent characteristics of biological medicinal products, including structural and functional heterogeneity, strong dependence of quality attributes on manufacturing process parameters and starting materials, and limited analytical accessibility of certain quality aspects, preclude exhaustive control and necessitate a differentiated analytical control approach proportionate to patient risk. The methodological framework of the proposed model is based on the concept of quality by design, in which the quality control strategy is constructed through a consistent logical sequence from the intended use of the medicinal product and its clinical context to the quality target product profile, identification of critical quality attributes, and definition of requirements for analytical methods and the stringency of their application. It is shown that critical quality attributes should be defined based on the potential impact of their variability on product safety and efficacy rather than on the availability of analytical methods, and should be classified according to the nature of their influence on the clinical profile.

The analytical control strategy is considered as a predesigned set of analytical and procedural controls aimed at managing quality risks by reducing the probability of analytical errors and increasing the capability to detect clinically and technologically significant changes in product quality. The article discusses the principles of analytical method fitness for purpose, the role of the analytical target profile, and mechanisms for differentiating analytical control stringency depending on the level of risk and the stage of the product lifecycle.

It is shown that the analytical control strategy represents a dynamic system that evolves as knowledge of the product and manufacturing process increases, shifting from broad exploratory objectives during early development to regulated release and stability testing during commercial manufacturing. The proposed approach provides a methodological basis for the design, justification, and controlled revision of analytical control strategies for biological medicinal products throughout their entire lifecycle.

This brief overview and lecture focuses on one of the key areas of regenerative medicine: tissue engineering using stem and multipotent cells. The technologies discussed include classical tissue engineering using scaffolds, spheroid approaches, and cell sheets. The article examines and highlights the basic principles of tissue engineering (the triad of components and key technologies) and the limitations associated with the use of this group of approaches in clinical practice.

Several hormones and paracrine factors like insulin, adiponectin, leptin, IGF-1, -2 activate phosphoinositide 3-kinase (PI3K) signaling pathway affecting adipogenic differentiation of multipotent mesenchymal stromal cells (MSCs). This study was aimed to develop an instrument for tracking a link between hormonal signaling and long-term cell response like cell differentiation using genetically encoded PH-GFP biosensor. Human Telomerase-Immortalized (hTERT) MSC was transduced by a lentiviral vector with the sequence based on pleckstrin homology domain of Bruton’s tyrosine kinase and GFP (PH-GFP). Then, the cell line was separated using flow cytometry. MSC^GFP-PH were examined for the PDGFand insulin-induced fluorescent responses, differentiation potential using standard differentiation protocols and proliferation rate applying neural networks for cell quantity counting. The obtained hTERT MSC^GFP-PH cell line retains intact adipogenic and osteogenic potential. It allows for the real-time detection of insulin and other PI3K-agonist signaling in living cells, both in the undifferentiated MSC state and during adipogenic differentiation. To simplify the detection of signaling events, we applied a computational approach to improve signal visualization and used a special metric to assess signaling activation. MSC^GFP-PH may be a useful instrument for investigating PI3K signaling in real time during long-term cellular processes such as adipogenic differentiation. This computational approach may represent a convenient option for detecting signaling by the PH-GFP biosensor at the single-cell level.

Persistent systemic inflammation is a critical component in the pathogenesis of age-associated diseases, particularly chronic heart failure (CHF). Numerous studies have demonstrated elevated inflammatory biomarkers in patients with CHF. One mechanism sustaining this chronic inflammation is the accumulation of senescent cells, which secrete a complex of bioactive factors known as the senescence-associated secretory phenotype (SASP). Senolytic therapy, which selectively eliminates senescent cells and reduces associated inflammation, represents a promising therapeutic approach for CHF. Dihydroquercetin, a compound with demonstrated senolytic activity in experimental models and a proven safety profile in clinical trials for other indications, was selected for the present study.

Objective: to evaluate the effects of different doses of dihydroquercetin on senescent cell biomarkers and the clinical status of patients with CHF.

Materials and methods. This is a randomized, comparative, double-blind clinical trial. The study will enroll 60 patients following an unplanned healthcare visit for symptoms and/or signs of congestion due to heart failure requiring hospitalization and/or intensification of diuretic therapy. Patients will be randomized in a 2:1 ratio (n = 40 in Group 1; n = 20 in Group 2) to receive either 100 mg/day or 30 mg/day of a liquid water-soluble form of dihydroquercetin (Taxifolin Aqua, “Advanced Technologies” LLC) as an adjunct to standard of care. Randomization will be performed using randomization tables, assuming group homogeneity across baseline parameters. Recruitment is scheduled over a 9-month period at a single study center. The follow-up period will be 16 weeks from enrollment. Follow-up visits are scheduled at 8 and 16 weeks to assess quality of life, functional status, and senescent cell burden biomarkers.

Current status. Patient recruitment is currently ongoing.