Olga S. Prokopyuk

Influence of Factors of Cryopreservation and Hypothermic Storage on Survival and Functional Parameters of Multipotent Stromal Cells of Placental Origin

Human placenta is a highly perspective source of multipotent stromal cells (MSCs) both for the purposes of patient specific auto-banking and allogeneic application in regenerative medicine. Implementation of new GMP standards into clinical practice enforces the search for relevant methods of cryopreservation and short-term hypothermic storage of placental MSCs. In this paper we analyze the effect of different temperature regimes and individual components of cryoprotective media on viability, metabolic and culture properties of placental MSCs. We demonstrate (I) the possibility of short-term hypothermic storage of these cells; (II) determine DMSO and propanediol as the most appropriate cryoprotective agents; (III) show the possibility of application of volume expanders (plasma substituting solutions based on dextran or polyvinylpyrrolidone); (IV) reveal the priority of ionic composition over the serum content in cryopreservation media; (V) determine a cooling rate of 1°C/min down to -40°C followed by immersion into liquid nitrogen as the optimal cryopreservation regime for this type of cells. This study demonstrates perspectives for creation of new defined cryopreservation methods towards GMP standards.

Towards biobanking technologies for natural and bioengineered multicellular placental constructs

Clinical application of a large variety of biomaterials is limited by the imperfections in storage technology. Perspective approaches utilizing low-temperature storage are especially challenging for multicellular structures, such as tissues, organs, and bioengineered constructs. Placenta, as a temporary organ, is a widely available unique biological material, being among the most promising sources of various cells and tissues for clinical and experimental use in regenerative medicine and tissue engineering. The aim of this study was to analyse the mechanisms of cryoinjuries in different placental tissues and bioengineered constructs as well as to support the viability after low temperature storage, which would contribute to development of efficient biobanking technologies. This study shows that specificity of cryodamage depends on the structure of the studied object, intercellular bonds, as well as interaction of its components with cryoprotective agents. Remarkably, it was possible to efficiently isolate cells after thawing from all of the studied tissues. While the outcome was lower in comparison to the native non-frozen samples, the phenotype and expression levels of pluripotency genes remained unaffected. Further progress in eliminating of recrystallization processes during thawing would significantly improve biobanking technologies for multicellular constructs and tissues.

Effects of Implantation of Cryopreserved Placental Explants on the Behavioral Indices and Morphological Characteristics of the Cerebral Structures in Senescent Mice

The effects of implantation of cryopreserved placental explants (CPEs) on the behavioral phenomena were studied in adult young (6 months old) and in senescent (presenile ontogenetic period, 12 months old) mice; morphological characteristics of the cerebral structures were also examined in these animals. Implantation of CPEs significantly influenced the behavioral indices and adaptational capacities of senescent mice; the direction of such effects was sex-dependent. In male mice, manifestations of deadaptation (decreases in the mobility and exploration habits and an increase in anxiety) were observed, while in females the behavioral manifestations were opposite (positive). Implantation of CPEs led to partial smoothing of negative shifts in the morphological characteristics in the motor cortex and hippocampus of senescent mice. Therefore, CPEs can serve as a source of natural compounds and cell elements providing the activation of neurogenesis, formation of new neurons, and their proliferation in key structures of the brain. In prospect, CPE-based therapeutic agents can be used for the correction of age-related dysfunctions of the CNS.

Аналіз методологічних підходів при моделюванні гестаційного антифосфоліпідного синдрому

Experimental study of obstetric antiphospholipid syndrome may reduce morbidity and deaths of women and newborns. Aim. The aim of the work is to examine relevant literature on modeling obstetric antiphospholipid syndrome, to choose the most appropriate model and to reproduce it in accordance with existing clinical criteria. The current views on the modeling of disease depending on the purposes, selection of laboratory animals, treatment and dosing medications, evaluation of the effectiveness of simulation and experimental treatment were comprehensively analyzed. Results. According to the authors the most appropriate model is a model of active immunization of mice with the cardiolipin antigen, which was reproduced in the authors modification.