[Human subcutaneous adipose tissue subjected to cold shock as a source of viable cellular population with characteristics of multipotent mesenchymal stromal cells].

Cellular population with characteristics of multipotent mesenchymal stromal cells (MMSCs) was isolated from subcutaneous adipose tissue frozen without any cryoprotectant at -70 degrees C. Under critical for the adipose tissue condition, the cells retained their viability in vitro and ability of adhesion to plastic. Cellular population was homogeneous and represented by small cells (d - 7 microm) with fibroblast-like morphology. Cells were positively stained with Abs for the Abs: CD29, CD44, CD49a, b, d, CD73, CD90, CD105, CD166, HLA ABC. Cells were negative for CD34, CD45--markers of hematopoietic cells, CD31--marker of endothelial cells, Stro-1, as well as for HLA DR, DP, DQ (flow cytometer analysis). Being induced to differentiate in vitro, the cells were able to differentiate into cells similar to cells of bone, adipose and cartilage tissue. Karyological assay of the cells isolated from human adipose tissue subjected to cold shock revealed diploid set of chromosomes, 46, XX, without aneuploidy and structural reconstructions of chromosomes. Thus, it has been established that, under extreme condition for the organism, the population of cells with a phenotype similar to miltipotent mesenchymal stromal cells is preserved in subcutaneous adipose tissue.

[Differentiation of multipotent mesenchymal stromal cells of bone marrow into cells of cartilage tissue by culturing in three-demential OPLA scaffolds].

Bone marrow multipotent mesenchymal stromal cells represent a perspective material for engineering of human three-dimensional transplants of cartilage tissue. We are demonstrated the opportunity of the directed differentiation of BM MMSC in cells of cartilage tissue by culturing them in three-dimensional scaffolds, presented by polymer OPLA in medium with inductors of chondrogenesis. For loading cells in porous scaffolds used method which essence consist in saturation of polymeric blocks by cellular suspension with the subsequent centrifugal force of cells in scaffolds and culturing of engineering constructs for 28 days in chondrogenic medium. Histological analysis derived in vitro of three-dimensional transplants showed uniform distribution of cells in the matrix with morphologically distinct chondrocytes-like cells of hyaline cartilage. Immunohistochemical analysis detected aggrecan and collagen type II within the extracellular matrix. Preclinical the researches lead on a livestock of immunodeficient mice have shown not toxicity of the engineering constructs.

[Cultivation and transplantation of boar type A spermatogonia].

A cell population enriched with type A spermatogonia has been isolated from the boar testes. Cell types occurring during isolation were morphologically characterized, factors maintaining the cultured spermatogonia in the undifferentiated state were studied, and these cells were transferred to sterile recipients preliminarily treated with busulfan. The cells of spermatogenic epithelium cultivated in vitro for 24 h were used for transfer experiments. The transfer efficiency was estimated within 27 and 29 days according to the histological picture of the testes and the isolated cultures. Spermatogenic cells at various developmental stages and a few Sertoli ells and spermatozoa were found on sections and in cell suspensions. Sperm samples could be taken from recipient boars within nine months after the transfer. Microsatellite analysis of DNA showed the endogenous pattern of spermatogenesis. Thus, it was shown that spermatogenic donor cells can restore and maintain spermatogenesis of a recipient for at least 30 days. However, the donor cells were fully forced by the recipient reserve cells, type A0 spermatogonia, within eight to nine months.

[Characteristics of human mesenchymal stem cells isolated from bone marrow and adipose tissue].

Populations of human mesenchymal stem cells were derived from bone marrow and adipose tissue. Here analysis of six individuals is represented. Cells were isolated, expanded and evaluated by the expression of surface antigens using flow cytometry. These cells displayed similar characteristics for many markers. Cells isolated from bone marrow and adipose tissue were found to be homogeneously positive for CD13, CD44, CD90, CD105, and negative for CD45, CD34, CD31 and CD117. Besides, differences in surface antigene CD10 expression between narrow and adipose tissue-derived cells were detected. All these findings indicate that both bone marrow and adipose tissue are important sources of mesenchymal stem cells, which could be used in cell therapy protocols.

[Comparative analysis of cell populations with a phenotype similar to that of mesenchymal stem cells derived from subcutaneous fat].

Two cell populations with a phenotype similar to that of mesenchymal stem cells (MSC) with different characteristics for expression of surface antigene CD34 were derived from subcutaneous fat. CD34-positive cells were derived from subcutaneous fat of the inferior eyelid obtained during transconjuctival blepharoplasty. CD34-negative cells were derived from adipose tissue obtained during lipoaspiration from the same patients. These cells displayed common characteristics for morphology and expression of basic markers characterizing them as mesenchymal stem cells. On being induced for differentiation, these two cell populations were able to differentiate to cells of adipose (adipocytes), bone (osteoblastes, osteocytes), cartilage (chondroblasts, chondrocytes), and nervous (neurons, astrocytes and oligodendrocytes) tissues.