Analysis of Reparative Activity of Platelet Lysate: Effect on Cell Monolayer Recovery In Vitro and Skin Wound Healing In Vivo.

Platelet lysate prepared from donor platelet concentrate and pooled according to a developed technique stimulates migration of multipotent mesenchymal stromal cells of the human adipose tissue and promotes healing of the monolayer defect in cultures of human fibroblasts and multipotent mesenchymal stromal cells in vitro in concentrations close those of fetal calf serum (5-10%). Lysate of platelets from platelet-rich rat blood plasma stimulated healing of the skin defect by promoting epithelialization and granulation tissue formation. The regenerative properties of platelet lysate in vivo increased with increasing its concentration.

Scleractinium coral aquaculture skeleton: a possible 3D scaffold for cell cultures and bone tissue engineering.

Cytocompatibility of 5 coral aquaculture skeleton species derived from two families (Acroporidae and Pocilloporidae) was studied over the course of in vitro culturing in continuous human fibroblast culture by the MMT test. Biocompatibility and capacity of scaffold to "transfer" cell cultures (specifically, multipotent mesenchymal stromal cells) to sites of implantation were studied in vivo by subcutaneous implantation of skeletal fragments to rats. All coral skeleton aquaculture specimens were cytocompatible (nontoxic and with surface matrix characteristics satisfactory for cells), biocompatible, and could be tried as 3D matrices for bone tissue engineering.

Human platelet lysate as a promising growth-stimulating additive for culturing of stem cells and other cell types.

We compared the composition and biological activity of fetal calf serum and platelet lysate from donor platelet concentrate. In platelet lysate, the concentrations of alkaline phosphatase, lactate dehydrogenase, creatinine, and mineral metabolism parameters were lower, while parameters of lipid and protein metabolism were higher than in fetal calf serum. The concentrations of growth factors (platelet-derived (AA, AB, BB), vascular endothelial, insulin-like, and transforming growth factor ß) in platelet lysate 1.7-148.7-fold surpassed the corresponding parameters in fetal calf serum. After replacement of fetal calf serum with platelet lysate in the culture medium (0, 25, 50, 75, and 100%), the count of multipotent mesenchymal stromal cells on day 7 (in comparison with day 1) increased by 154.8, 206.6, 228.2, 367.7, and 396.5%, respectively. Thus, platelet lysate can be an adequate non-xenogenic alternative for fetal calf serum.

Study of growth parameters and potentialities of differentiation of multipotent mesenchymal stromal cells from rat bone marrow in vitro.

The dynamics of growth and proliferative activity of the population of multipotent mesenchymal stromal cells from rat bone marrow was studied during 7 passages. The efficiency of colony formation, the morphology of multipotent mesenchymal stromal cells, and the possibility of spontaneous and induced differentiation were studied. The rat bone marrow fibroblast-like multipotent mesenchymal stromal cells are capable of clonal growth; their proliferative activity and the yield remained high until passage 4, but then decreased. Induction of osteo- or adipogenic differentiation of bone marrow multipotent mesenchymal stromal cells increased the percentage of morphologically modified cells carrying specific markers.

[Application of cell technologies to tissue defect closure in oncology].

In vivo and in vitro experiments on the application of cell technologies to tissue defect closure were conducted; autologic mesenchymal stem cells on 3-demensional matrices were used. The authors analyze the results of the application of bioengineering tissue equivalents for the closure of soft tissue and upper airway defects after extensive resections performed in 52 oncological patients. Tissue equivalents with stem cells provide engraftment and long-term graft functioning; they also modify wound surface, thus stimulating wound epithelization. In this study the application of tissue equivalents led to wound healing and functional recovery in 87% of patients.

Effects of various modes of sonication with low frequency ultrasound on in vitro survival of human tumor cells.

The effect of low frequency ultrasound in the cavitation (5-15 sec exposure) and subcavitation (5-210 sec exposure) modes on in vitro survival of cultured human tumor cells was studied. Analysis of the dose-effect curves and mathematical estimation of the effects low-frequency ultrasonication in the subcavitation mode on cell membranes helped to choose time intervals of tumor cell sonication ensuring potentiation of the cytostatic effects.

[The prediction of the individual chemosensitivity of human malignant tumors: the biological patterns and clinical results]. / Prognozirovanie individual'noi khimiochuvstvitel'nosti zlokachestvennykh opukholei cheloveka: biologicheskie zakonomernosti i klinicheskie rezul'taty.

Individual laboratory prediction of the sensitivity of human neoplasms to chemicals is justified on the basis of the authors' own data and those available in the literature. The potentialities and limitations of methods for individual prediction, as well as clinical results are discussed. Data are given on some biological mechanisms responsible for tumor growth which have been obtained in in vitro and in vivo tests with human tumor tissue and carcinoma cells. Further perspectives of this line of studies are discussed.

[A comparative evaluation of 2 methods for cloning human lung tumors]. / Sravnitel'naia otsenka dvukh sposobov klonirovaniia opukholei legkogo cheloveka.

Clonogenic capacity of tumor cells from 59 human lung cancer specimens was investigated in Petri dishes (24 specimens) and in capillaries (40 specimens), and parallel cloning was done in 5 cases. The results were as follows: (a) bacterial contamination in capillaries was noted less frequently than in Petri dishes (12.5 vs. 25%); (b) cells did not form colonies in capillaries in 42.5% and in Petri dishes in 25%; (c) colony-forming effectiveness in capillaries was 3.46-fold higher than that in Petri dishes; (d) the amount of tumor cells for analysis in capillaries is 10-fold less than that for analysis in Petri dishes. However, the absolute number of colonies, indispensable for assessment of tumor sensitivity to antitumor action (not less than 15 colonies per capillary or Petri dish) was obtained in 67% in Petri dishes and in 33% only in capillaries.

[Preparation of tumor cells from human lung cancer tissue for the purpose of cloning]. / Poluchenie opukholevykh kletok iz tkanei raka legkogo cheloveka s tsel'iu klonirovaniia.

Various approaches to isolation of tumor cells are analyzed on the basis of examination of 46 human pulmonary tumors of different histologic types. Mechanical treatment alone resulted in the death of the majority of tumor cells. Short trypsin treatment (for 10-15 min in several stages) proved to be the most suitable for disaggregation of small-cell carcinoma, adenocarcinoma, and carcinoid samples. A specific approach has been developed for isolation of cells from squamous-cell carcinoma, consisting in tumor treatment with 0.25 percent trypsin at 4 degrees C for 16-18 h, followed by DNAase, collagenase, and trypsin treatment, and then again trypsin treatment at 37 degrees C for 10-15 min. The suggested approaches permit a harvest of at least 6 x 10(6) tumor cells from 1 g of tissue, with more than 70 percent of these cells viable.

[Assessment of individual sensitivity to chemotherapy of human pulmonary tumors transplanted to subrenal capsule of CBA mice]. / Otsenka individual'noi chuvstvitel'nosti k khimiopreparatam opukholei legkogo cheloveka pri ikh transplantatsii pod kapsulu pochki myshei linii CBA.

The individual response to chemotherapy was investigated in 17 heterotransplants of human lung tumours of different histogenesis by the subcapsule test. It is established that the subcapsule test is of high informativity only when testing the individual resistance of lung tumours, especially adenocarcinomas, to cytostatics.

[Several parameters of growth of heterografts of human lung tumor in the subrenal capsule of CBA mice]. / Nekotorye parametry rosta geterotransplantatov opukholei legkogo cheloveka pod kapsuloi pochki myshei linii CBA.

28 species of human lung cancer of the III-IV stages were investigated. The maximal growth of heterotransplants was found for them with the initial area of 1.2-1.5 mm2. The analysis of the data obtained has shown that the samples of lung adenocarcinomas grow more slowly than those of squamous cell and small-cell carcinomas and also of carcinoid.

[The clonogenic capacity of human lung cancer cells]. / Klonogennaia sposobnost' kletok raka legkogo cheloveka.

Comparative studies of clonogenic ability of twenty four lung cancer samples have been performed in semisolid agar in vitro and in diffusion chambers in vivo. It is concluded that plating efficiency of lung cancer cells varies from 0.00002% to 0.03% in vitro assays and from 0.01% to 0.11% in vivo. A small number of colonies (less than 20) in 15 specimens from 16 does not permit using this method for preclinical individual sensitivity test. The cloning of lung cancer in vitro gives the efficiency more than twenty colonies per a dish in 5 cases of 11. There is some evidence that these models can be used for prediction of treatment response of individual tumors to chemotherapy.

[Evaluation of the proliferative activity of malignant human tumors by an indirect immunofluorescent method using antithymidine antibodies]. / Otsenka proliferativnoi aktivnosti nekotorykh zlokachestvennykh opukholei cheloveka nepriamym immunofluorestsentnym metodom s pomoshch'iu antitimidinovykh antitel.

155 samples of human malignant tumours were studied by the immunofluorescent method and the antithymidine antibodies index-labelling (IL--% of tumour cells in the S-phase). Wide individual variability (0.1-73%) of IL of the studied tumours was shown to be present and to be preserved inside the tumours which were homogeneous by the histological type and developmental stage. A correlation between the labelling index and tumour differentiation in the gastric adenocarcinoma is observed.

[T- and B-cell functional activity in the immune response in viral carcinogenesis]. / Issledovanie funktsional'noi aktivnosti T- i B-kletok v immunnom otvete pri virusnom kantserogeneze.

The functional activity of the spleen cells, their ability to produce antibody-forming cells on the model of the adoptive transfer, activity of T-helper-cells and B-precursor-cells of antibody producers in the tumour growth induced by monkey SA7 (C8) adenovirus were studied. The inoculation of the tumourigenic virus to the newborn mice CBA, inhibit antibody formation in the system of adoptive transfer already in the latent period with a sharp inhibition at the terminal stages of carcinogenesis. The helper activity is suppressed earlier than the activity of B-precursors of antibody producers. The obtained data indicate a considerable immunodepressive action of monkey adenovirus SA7 (C8).

[Changes in the activity of oxidative enzymes in thymus cells of CBA mice during carcinogenesis induced by simian adenovirus SA7(C8)]. / Izmenenie aktivnosti okislitel'nykh fermentov v kletkakh timusa myshei CBA v protsesse kantserogeneza, indutsirovannogo obez'ian'im adenovirusom SA7(C8).

Microspectrophotometry was used to study the activity of oxidative enzymes in the cortical and medullar regions of the thymus, in epithelial stromal cells of CBA mice at varying time of postnatal development and during induced virus cancerogenesis. It was shown that in the thymus of control animals, the activity of NAD-, NADP-diaphorases and beta OBDH reached maximum on the 30th day of postnatal development. The animals exposed to virus manifested a distinct lowering of the activity of the enzymes even in the latent period of cancerogenesis. Variations of the activity of the test enzymes during postnatal development of control animals and at varying time of cancerogenesis appear to be related to functional potencies of the thymus.

[Mechanisms of changes in the mass of the organs central to immunity during adenovirus carcinogenesis]. / O mekhanizmakh izmeneniia massy tsentral'nykh organov immuniteta pri adenovirusnom kantserogeneze.

The spleen cells transfer from mice CBA at the 25th day of the carcinogenesis latent period induced by adenovirus SA7(S8) to newborn syngeneic animals caused the graft versus host reaction in them. There was splenomegaly and progressive decrease in weight of the recipients' thymus. Analogous alterations of lymphoid organs were noted in the animals infected during the neonatal period by oncogenic adenovirus SA7(C8). Results showed that adenoviral carcinogenesis had some manifestations of autoimmune disease.