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Vol 9, No 2 (2014)

Articles

Professor Alexander Maximow: evolution of the ideas

Deev R.V.

Abstract

Russian scientist-histologist professor of the Military Medical Academy Alexander Alexandrovich Maximow is one of the founders of conception about stem cells in general, and hematopoietic stem cells, particularly. Started to study the morphology of inflammation, within three decades he explored in detail the hemocytopoiesis in bone, described the different stages of this process. He formulated the idea of «polyblasts», a pluripotent precursor with lymphocyte phenotype (rounded cells with spherical nucleus), which moved out of vascular bed and were able to differentiate both into wandering phagocytic cells and fibroblasts in the inflammation site. The study of hematopoiesis in the evolutionary series of animals from cartilaginous fish to mammals led to the dominance of the hematopoiesis unitary model in embryogenesis and postnatal organisms' life. Despite the serious scientific debates around the world in the future (60-ies of XX century), exactly this model of hematopoiesis has been proved by the method of radiation chimeras and became accepted as a theoretical basis for hematopoietic stem cell transplantation.
Genes & Cells. 2014;9(2):6-14
pages 6-14 views

UNIQUE BETA-LIKE DNA POLYMERASE FROM ХРОМАТИН OF HUMAN ACUTE MYELOID LEUKEMIA HL-60 CELLS

Buchvostov A.A., Orlov A.P., Shatalov O.A., Kusnetsov D.A.

Abstract

Human acute myeloid leukemia cells HL-60 over expresses a beta-like DNA polymerase (EC 2.7.7.7) which is found to be a chromatin associated single subunit protein (66.5 kDa) purified by original extraction/gel filtration procedure allowing to gain the 122,000-fold purification degree as corrected to a total cell protein. The enzyme possesses some key DNA pol р-specific catalytic properties such as the processing of short (200-250 n) single strand DNA sequences, activation in the presence of 200 mMKCl, resistance to N-ethyl-melamide and Aphidicolin, lack of 3',5'-exonuclease activity. A possible significance of the unique enzyme studied as a target for its pharmaceutical inhibitors is under discussion.
Genes & Cells. 2014;9(2):46-52
pages 46-52 views

Optimization of conditions of skin and gingival mucosa derived human fibroblasts obtainment and cultivation

Zorin V.L., Kopnin P.B., Zorina A.I., Eremin I.I., Lazareva N.L., Chauzova T.S., Samchuk D.P., Petrikina A.P., Eremin P.S., Korsakov I.N., Grinakovskaya O.S., Solovieva E.V., Kotenko K.V., Pulin A.A.

Abstract

For optimization of conditions of skin and gingival mucosa derived human fibroblasts obtainment and cultivation in this work we performed comparative analysis of 11 fetal bovine serums (from different manufacturers) and autologous human serum. Our analysis included also comparison of 3 serum-free media and 4 serum-reduced media. Evaluation criterion at the step of primary cell culture was colony-formation efficiency and during subsequent cultivation - fibroblasts population doubling time. Results revealed that ability of skin-derived fibroblasts to form colonies could vary 4 and more times and gingival mucosa derived fibroblasts - 1,5-1,8 times depending on used for cultivation type of serum or medium. Clonal analysis showed that fibroblasts in culture form 3 types of colonies consisting of different amount of cells and different phenotype. Ratio of colonies could be variable and depend on type of used medium. It was noticed during cultivation that addition of different sera to standard medium (DMEM) led to changing of cell's doubling time. In general, successful obtainment of fibroblasts primary culture determined by selection of appropriate serum and medium providing development of large colonies of rapidly proliferating cells. It was noticed during cultivation that addition of different sera to standard medium (DMEM) led to changes in cell's doubling time.
Genes & Cells. 2014;9(2):53-60
pages 53-60 views

Proliferative and differentiation potential of adipose tissue-derived mesenchymal stromal cells in the presence of platelet lysate

Rogulska O.Y., Revenko O.Б., Petrenko Y.A., Petrenko A.Y.

Abstract

Clinical application of mesenchymal stromal cells requires the development of serum-free culture medium for optimal cell growth and differentiation. Platelet lysate as an alternative for xenogeneic fetal serum is able to promote expansion of human adipose tissue-derived mesenchymal stromal cells. Platelet lysate stimulates the proliferative activity of cells and increases the efficiency of colony formation. When cultured mesenchymal stromal cells in the presence of platelet lysate we identified colonies of the same types as in the presence of fetal bovine serum, but their ratio changes in favor of dense and mixed colonies compared with diffuse colonies. Size and density of the cell colonies formed in the presence of platelet lysate are higher than in those in serum. After expansion in medium with 10% platelet lysate mesenchymal stromal cells retain the ability to differentiate into osteogenic and adipogenic directions while efficiency of osteogenic differentiation of these cells is better than of cells previously expanded in the presence of serum. The results of current study show that platelet lysate is a promising natural substitute for xenogeneic serum in optimizing culture technology of human mesenchymal stromal cells for regenerative medicine and tissue engineering.
Genes & Cells. 2014;9(2):63-67
pages 63-67 views

Hyaluronan-based bioplastic material as a scaffold for biomedical cell-based express-product for skin regeneration

Kalmykova N.V., Spichkina O.G., Ellinidi V.N., Rakhmatullin R.R., Moiseev S.I.

Abstract

This study is designed to investigate the behavior of human fibroblasts and keratinocytes on hyaluronan-based bioplastic material «G-DERM». Cell viability, proliferation, interleikin-6 and keratinocytes growth factor secretion were investigated. The results suggested that «G-DERM» was not toxic and supported cell adhesion and spreading. Histological analysis showed that «G-DERM» properties and porous structure provided distribution of skin cells by cell-type manner. Fibroblasts on «G-DERM», being quiescent, kept their synthetic activity and produced tissue-specific factors of wound healing. According to study results «G-DERM» can be used as 3D cell matrix for regenerative medicine.
Genes & Cells. 2014;9(2):68-75
pages 68-75 views

Morphologic assessment of compensatory abilities of scalp skin cells in staged dermotensia

Checmareva I.A., Paklina O.V., Sharobaro V.I., Vaganova N.A., Gordienko E.N., Vaganov N.V., Grechishnikov M.I.

Abstract

The purpose of the research was morphologic (histologic and electron-microscopic) study of compensatory adaptational reaction of skin of the scalp after staged expansion. Morphologic assessment of adequacy of the expansion and compensatory abilities of cells of tissues under expansion was carried out in 27 patients. It was ascertained in the course of the work that the secondary dermotensia (twofold, threefold, fourfold) did not provoke degenerative changes in expanded scalp tissues, ultrastructural changes of cells of epidermis and dermis characterized adaptable reactions that did not exceed their compensatory abilities.
Genes & Cells. 2014;9(2):76-79
pages 76-79 views

Decellularized human umbilical arteries retain their mechanical properties

Nasredinov A.S., Lavreshin A.V., Lebedeva E.A., Anisimov S.V., Vavilov V.N., Kurapeev D.I.

Abstract

Tissue engineered vascular grafts can fulfill a clinical need in biological prostheses in reconstructive cardiovascular surgery. Decellularized arteries do not cause immune response, are biocompatible, could be reseeded with recipient cells und thus are attractive scaffolds for vascular tissue engineering. Earlier we developed a decullarization method for human umbilical arteries and proved its effectiveness morphologically. The purpose of this study was to evaluate mechanical properties of the decellurized human umbilical arteries, also after long-term storage. 3 groups of vessels were investigated: I group - native arteries, II group - decellularized arteries, III group - decellularized arteries, stored in phosphate buffered saline for 10 months. Samples were stretched until rupture on the Instron universal testing machine; herewith strain and stress were recorded. The same way the suture retention strength was estimated. Also burst-pressure, that characterized the total strength of the samples, was investigated. Intergroup differences in maximum strain both longitudinal and transverse directions, suture retention strength and burst pressure were not significant. Decellularized human umbilical arteries retain their mechanical properties, and that indirectly confirms extracellular matrix preservation. Thus they are attractive source for small diameter tissue engineered vascular grafts.
Genes & Cells. 2014;9(2):80-86
pages 80-86 views

Differentiation of epitheliocytes of mesonephric and paramesonephric ducts in human embryogenesis

Zheglova M.Y.

Abstract

The aim of the research was to give the morphofunctional characteristics of embryonic histogenesis of the female genital tract abortuses. We have studied 50 abortuses at the age of 4-12 weeks of gestation. The fixation of material and of the sample preparations were carried out according to the principles of quantitative histochemistry. According to the data obtained, paramesonefric ducts are found in the composition of the urogenital blastema to which coelomic epithelium cells can migrate. Since in this time the more differentiated ducts are mesonefric and archinephron is the organ that carries out its function, it is undeniable that the development of paramesonefric ducts goes on vector of the cranio-caudal growth of mesonefric ducts. The formation of a common utero-vaginal canal occurs through the convergence of the paramesonefric duct walls. Herewith the septum between ducts is persisted. Epitheliocytes in the medial walls of the ducts are deformed, nuclei decrease in size. The shape of the nuclei changes and rectangular nuclei dominate. The cells which have a light nucleoplasm are detected, in other cells there is a piknotic nucleus in the center; there are cells with nuclei having the semilunar shape, cells with two fragments of the nucleus, one of which is dark and the other one is light; nucleus with cuspidal protrusions at one pole, cigar-shaped nucleus. The formation of the complex relationships between the paramesonefric ducts at the entrance into mesenchymal block allows to make a suggestion on the critical period of the female genital tract development.
Genes & Cells. 2014;9(2):87-92
pages 87-92 views

Histotechnical solutions for quality improvement of nucleic acid specimens extracted from paraffin blocks

Vaganova A.N.

Abstract

With the growing body of information about molecular nature of a set of diseases, the question of molecular biology techniques acceptability for histological material study came up. Currently available pathology archives could provide vast quantity of material for retrospective studies, however, the most frequently used fixative formalin dramatically damages nucleic acids in the tissue. A similar problem arises with the material that come into the pathology department and undergoing conventional histological processing in nowadays. In recent years with the entry into the practice of diagnostic methods based on molecular biology methods it has become especially important to obtain high-quality samples of nucleic acids. Research devoted to development of new approaches of histological material handling from one side and solutions for nucleic acids extraction from tissue after standard histological treatment on the other side revealed a number of ways to overcome the problem of poor quality of nucleic acids isolated from the paraffin block. The aim of this review is to highlight the modifications histological processing techniques that allow prepare nucleic acids samples of higher quality.
Genes & Cells. 2014;9(2):96-101
pages 96-101 views

Related allogeneic cord blood and bone marrow hematopoietic stem cell transplantation for Fanconi anemia: a case report

Shul'ga S.Y., Boyarchuk N.G., Potapov I.V., Skorobogatova E.V., Dyshlevaya Z.M., Donyush E.K., Prihodko A.V., Isaev A.A.

Abstract

Successful cord blood and bone marrow hematopoietic stem cell transplantation for Fanconi anemia described. The feature of this clinical case is using cord blood unit from private cord blood bank. Private cord blood banks in the future can make a significant contribution to the structure of the autologous and allogeneic transplantations in oncohematology.
Genes & Cells. 2014;9(2):102-103
pages 102-103 views

Replacement of the anterior-lateral wall of the maxillary sinus with allografts

Shcherbakov D.A., Lebedeva A.I., Karimova A.I.

Abstract

The aim of the work was a comparative experimental and clinical study of efficacy of the bone and cartilage allografts in restoration of defects in the anterior-lateral wall of the maxillary sinus. Experimental studies were performed on rabbits Chinchilla. After modeling the defect of the anterior wall of the maxillary sinus the grafting was performed with cartilage allografts in the first experimental group (n = 12), in the second group (n = 12) - bone allograft was used as a biomaterial. In the clinical part of the study we evaluated the effectiveness of treatment in patients after maxillotomy with the reconstruction of the anterior-lateral wall with the cartilage (n = 21) or bone (n = 24) allografts. We found that both types of allografts induced reparative osteogenesis in the field of bone defect. Some features in regeneration process depending on the type of biomaterial were shown. Clinical studies demonstrated the efficacy of the cartilage and the bone allografts in replacement of the anterior-lateral wall of the maxillary sinus. In all cases a positive clinical outcome, the integrity of the anterior-lateral wall of the sinus, was achieved. Both allografts were replaced by lamellar bone tissue and that's why could be considered as an alternative of autogenous bone transplants in such operations. In patients with dolichocephalic skull and deep canine fossa it is appropriate to use the cartilage allografts characterized by higher elastic properties.
Genes & Cells. 2014;9(2):104-109
pages 104-109 views

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