Investigation of stem cells differentiation substantiates the reasonability of use and confirms clinical effectiveness of new developments in cellular technology. Immunocytochemical methods of revealing of synthesized marker proteins are widely used in evaluation of stem cells differentiation. This review contains characteristics of the most often used markers of neural differentiation that are used in investigation of histoblastic potencies of stem [including neural stem/progenitor) cells. These neuromarkers include nuclear protein of nerve cells NeuN, beta-tubulin III, protein MAP2 associated with microtubules, proteins of neurofilaments, neuron-specific enolase, synaptophysin and molecule of nerve cells adhesion CPSA-NCAM] connected to polysialic acid. The study evaluates the specificity of these markers and reveals the advantages and disadvantages of methods used for their detection. It is emphasized that detection of some proteins expression cannot serve as an unequivocal evidence of stem cells differentiation into neurons. The most neuronspecific markers are: NeuN, neurofilament proteins and synaptophysin.
We investigated affect of non-ionic block copolymer (Pluronic) P85 on in vitro proliferation and osteogenic differentiation of human mesenchymal stem cells (MSCs). We demonstrated that Pluronic P85 at final concentration 0,1-1% was cytotoxic to MSCs. Addition of Pluronic P85 at final concentration 0,001-0,01% increased proliferation of MSCs. Pre-incubation of MSCs with Pluronic P85 increased osteogenic differentiation efficiency of cells.
Here we preset a method for tissue engineering construct based on highly porous PTFE with nanostructured multifunctional biocompatible non-degradable coating (MBNC) with composition of Ti-Ca-P-C-O-N. It is shown that scaffolds composed of PTFE with MBNC are suitable for adipose-derived multipotent stromal cells CADMSJ active adhesion and proliferation. Under osteogenic stimulation ADMS cells were differentiated to osteogenic lineage, which was confirmed by positive immunohistochemical reaction for osteopontin and osteonectin. Using the experimental model of critical calvarial defects in rabbits we investigated the influence of tissue engineering construct [membrane from highly porous PTFE with MBNC carrying ADMS cells) transplantation on bone defect regeneration. It was shown that under tissue engineering construct the bone defect was completely closed with newly formed bone tissue in 3 and В months after transplantation.
New efficient methods for separation and cryopreservation of nucleated cells from umbilical cord blood, which allow you to save a quantitative and qualitative composition of nucleated cells (including hematopoietic stem] after discharge, and demonstrate a high level of preservation of structural and functional properties and viability of CD45+ and CD34+-cells after cryopreservation.
In the research there has been estimated an immune correcting activity of cryopreserved and native fetal liver cells (FLCs) of different gestation terms in experimental model of local GVHR CIGVHR). IGVHR was induced in C57BI/6 mice by means of subcutaneous introduction of CBA/H lymph nodes CLN) cells into the hindpaw pad. In 24 hrs after initiation of IGVHR the mcie were intravenously injected with either native CnFLCs) or cryopreserved FLCs (cFLCs) of the 14th and 18th gestation days of CBA mice. To the 5th day after initiation of pathology the following indices were estimated: GVHR index, content of T (FoxP3+, CD4+CD25+ cells) of LNs with flow cytometer FACS Calibur (Becton Dicknson, USA), the expression of tfg-beta gene by means of PCR; content of IL-2, IL-10, TNF-alpha in blood serum of animals by means of immune enzyme method with analyzer Stat Fax 2100 (USA). It has been established that during induction of IGVDR there are manifested the signs characteristics for the pathologies of immune genesis. Under development of GCH, FLCs minimize clinical signs of pathologies and intensity of development of immune inflammatory reaction. Therapeutic activity of FLCs in greater extent correlated to the content in LNs of F0XP3+ cells and expression of tgf-beta gene, but not to the content of CD4+CD25+ cells and rate of their fluorescence. With the increasing of gestation terms from 14 post-coital days to 18 ones immune correcting activity of FLCs reduced. However after cryopreservation the FLCs-18 gained immune correcting activity of nFLC-14.
The questions concerning condition and prospects of regional development of the cellular technologies are discussed in the work. The topicality of support of scientific researches in the field of biomedical technologies from outside the state structures, private enterprises, investors and venture funds is underlined. The concrete examples of such interaction are demonstrated and the reasons of their possible insufficient efficiency are analyzed. The information about development of the cellular technologies in the Ural Federal district is resulted. Possibility of advancement of new biomedical technologies in the regions at the expense of budgetary funds within the limits of existing regional target programs (priority directions] and theirs commercial realization by creation of small enterprises and/or the private-state partnership by means of target programs on development of innovative activity is shown.