Biomedicinskie kletochnye produkty (BMKP) — dermal'nyj ekvivalent kozhi (DEK) i biologicheskij ekvivalent kozhi (BEK) — eto allogennye ekvivalenty dermy i kozhi cheloveka, primenyaemye dlya lecheniya v sluchayah kriticheskoj poteri kozhi. Odnim iz vazhnyh voprosov razrabotki BMKP yavlyaetsya podbor uslovij hraneniya, sposobstvuyushchih uvelicheniyu sroka ih godnosti dlya rasshireniya vozmozhnostej logistiki i ispol'zovaniya. Cel'yu issledovaniya bylo opredelit' srok godnosti DEK i BEK putem ocenki morfologii i zhiznesposobnosti kletok v ih sostave i urovnej sekrecii imi faktora rosta endoteliya sosudov (VEGF) v processe hraneniya v fiziologicheskom rastvore pri komnatnoj temperature. Ispol'zuya MTT-test i okrasku vital'nym krasitelem, my ustanovili, chto pri hranenii oboih vidov BMKP v fiziologicheskom rastvore snizhenie zhiznesposobnosti i metabolicheskoj aktivnosti kletok bolee chem na 50% proiskhodilo k 3–4 sutkam hraneniya, prichem v DEK bystree, chem v BEK. Morfologiya kletok, vydelennyh iz DEK i BEK posle 3 sutok hraneniya, ostavalas' neizmennoj. Posle pomeshcheniya BMKP v fiziologicheskij rastvor mezenhimnye stvolovye kletki v ih sostave prodolzhali sintezirovat' VEGF (pokazano metodom IFA), chto mozhet svidetel'stvovat' o sohranenii imi neobhodimoj sekretornoj aktivnosti.

The paper deals with an electrophysical device for combined electroporative-iontophoretic effect on biological membranes. The experimental results on the influence of a pulsed current source with displacement on the viability of living tissue equivalents (LTE) cells are presented. There were obtained values ​​of pulse electrophysical parameters providing preservation of LTE cell vitality. According to the obtained data we suggest that such electrophysical parameters can be used for drugs delivery through the equivalent of the round window membrane and for creation in the future a technology for the administration of drugs through biological barriers with the possibility of their penetration into the inner ear using pulsed electrical installations.

PURPOSE:To study of the effectiveness of the drug Melphalan as an antiproliferative agent during experimental proliferative vitreoretinopathy (PVR).MATERIAL AND METHODS:The experimental study used data from 24 eyes of 12 Chinchilla rabbits weighing 2.5-3.0 kg, which had PVR modeled in both eyes by intravitreal injection of a culture of heterogeneous activated fibroblast cells consisting of 200,000 cells in 0.1 ml. Treatment of experimental PVR was performed 1 day after the modeling process. In the first group of animals (6 eyes), 0.02 mg of Melphalan was administered intravitreally. In the second group of animals (6 eyes), 0.005 mg of Melphalan concentrated in 0.1 ml was administered intravitreally. Left eyes in both groups remained without treatment. Animals were observed for 1 month using biomicroscopy and ophthalmoscopy. 30 days after the animals were removed from the experiment, the eyes were enucleated, fixed in 10% buffered formalin and subjected to standard histological examination. The study of paraffin sections of the eyes was performed using the microscopic system «Leica» (Leica Microsystems, Germany) with built-in digital camera at the magnification of 200-600.RESULTS:In groups 1 and 2 of the study in the eyes of rabbits that received treatment, PVR was absent, unlike the eyes without treatment, where PVR remained. In group 1, where the dose of Melphalan was 0.02 mg in 0.1 ml, there were changes in the RPE (retinal pigment epithelium), which was regarded as a retinotoxic effect. Glial degeneration and thinning of the retina with disappearance of the photoreceptor layer (the outer nuclear and plexiform layers) resulted from the disturbance of retinal metabolism caused by RPE destruction. In group 2, structure of the retina remained more intact: isolated foci were noted with a decrease in the volume of the outer nuclear layer, shortening of rods and cones with preservation of the inner layers of the retina.CONCLUSION:A single intravitreal injection of 0.005 mg Melphalan had a positive therapeutic antiproliferative effect on the PVR model with minimal retinotoxic changes.

Сахарный диабет и его осложнения представляют собой самую распространенную эндокринную патологию в большинстве стран мира. Изучение патогенетических механизмов возникновения и прогрессии этого заболевания, а также поиск новых терапевтических средств и способов терапии остаются актуальными на сегодняшний день. Важнейшее значение для изучения сахарного диабета имеют экспериментальные модели. В обзоре обобщены данные о наиболее часто используемых животных моделях. Проанализированы и обсуждены механизмы стрептозотоцинового диабета как наиболее адекватной и легко воспроизводимой экспериментальной модели диабета. Рассмотрены значимые преимущества и недостатки описанных моделей.

In this review, we tried to elucidate the origin and development of different animal and human cell culture methodologies used to evaluate the effects of various factors and substances in vitro. Organ cultures and conventional two-dimensional cultures of dissociated cells of various types, such as primary, tumor, induced pluripotent, stem, and etc. have their advantages and drawbacks but usually do not represent accurate models for studying biological processes that take place in living organisms. Nowadays high-throughput cell assays on the basis of various methods of signal detection (optical utilizing colorimetric, luminescent and fluorescent methods of detection and electrochemical) are widely used at early stages of drug development for selection of the most active compounds and evaluation of their cytotoxic effects. The use of animals as models for drug testing is being criticized because of the lack of correlation between the results obtained in studies on them and on humans, and also because of the high cost and ethical issues. Therefore, much effort is put to create models based on human cells. This is how cultures emerged that utilize a three-dimensional network to simulate the architecture of tissues in vivo, and then so-called “organs-on-chips” – microfluidic microdevices combining several types of cells, that replicate physical and chemical parameters of the microenvironment of cells in living organisms. In summary, experimental cell models have come a long way from the whole organs cultivated in a growth medium to almost complete reconstruction of organs in vitro based on the cutting-edge engineering approach with the use of different cell types. This currently enables to replicate complex biological processes and study the influence of different substances and factors on them more successfully.

Изучение фенотипической пластичности и дифференцировочных способностей клеток является важной задачей клеточной биологии. В представленной работе исследована способность клеток поднижнечелюстной слюнной железы мыши к дифференцировке в гепатоцитарном направлении, а также проанализировано влияние вальпроевой кислоты, ингибитора гистон-деацетилаз, на эффективность дифференцировки. Показано, что после дифференцировки в клетках слюнной железы возрастает экспрессия генов гепатоцитарных маркеров (Aat, Afp, G6p, Pepck, Tat, Cyp3a13) и факторов транскрипции (Hnf-3α, Hnf-3β, Hnf-4α, Hnf-6). Вальпроевая кислота увеличивает специфичность гепатоцитарной дифференцировки, снижая экспрессию протоковых (Krt19, Hhex1, Cyp7a1) и ацинарных (Ptf1a) маркеров. После воздействия вальпроевой кислоты усиливается также эффективность гепатоцитарной дифференцировки, о чем говорит повышение уровня Alb и Tdo2 и увеличение секреции мочевины дифференцированными клетками. Изменений в характере метилирования промоторных областей генов не обнаружено, однако обработка вальпроевой кислотой и гепатоцитарная дифференцировка в значительной степени затрагивают метилирование гистона H3, ассоциированного с генами, ключевыми для гепатоцитарной дифференцировки. Таким образом, клетки поднижнечелюстной слюнной железы мыши способны к дифференцировке в гепатоцитарном направлении, а вальпроевая кислота увеличивает специфичность и эффективность их дифференцировки.