Yu. V. Gerasimov

Powers of differentiation of clonal strains of bone marrow fibroblasts

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Transfer of bone-marrow microenvironment by clones of stromal mechanocytes

Colonies consistign of clones of bone-marrow stromal fibroblasts, grown in monolayer cultures of mouse and guinea pig bone-marrow cells, transfer the hematopoietic microenvironment on retransplantation into the animal. Individual clones simultaneously form bone tissue and create a microenvironment for all three branches of medullary hematopoiesis: erythroid, myeloid, and megakaryocytio.

Characteristics of human bone marrow mesenchymal stem cells isolated by immunomagnetic selection

Immunophenotype of human bone marrow mesenchymal stem cells was studied after several culturing passages and after cryopreservation. Immunocytochemical analysis showed that bone marrow mesenchymal stem cells acquired homogeneity during in vitro culturing, but initially contained heterogeneous populations.

Effect of BMP-2 protein on the count and osteogenic properties of multipotent stromal cells and expression of cytokine genes in primary cultures of bone marrow and spleen cells from CBA mice immunized with bacterial antigens

We studied the effect of BMP-2 added to the culture medium on osteogenic and proliferative properties of multipotent stromal cells (MSC) and on the expression of cytokine genes induced by immunization of experimental animals with bacterial antigens. It is shown that the presence of BMP-2 in the culture medium stimulates proliferation of bone marrow MSC and especially spleen MSC (which was seen from enlargement of MSC colonies); improves the efficiency of MSC cloning; increases osteogenic activity of mouse bone marrow MSC; induces osteogenic differentiation of splenic MSC (osteogenesis is normally not observed in the spleen); reduces the number of macrophages in cultures; inhibits synthesis of mRNA for proinflammatory cytokines (IL-1β, IL-6, IL-8, TNF-α) that typically occurs in cultures of the bone marrow and spleen from animals immunized with S. typhimurium or group A streptococcus antigens. Bearing in mind that proinflammatory cytokines negatively affect osteogenic activity of the bone marrow, we can hypothesize that BMP-2 not only stimulates osteogenesis, but also provides optimal conditions for its realization by suppressing the expression of genes encoding these cytokines.

Proline-rich hypothalamic polypeptide has opposite effects on the proliferation of human normal bone marrow stromal cells and human giant-cell tumour stromal cells.

In the present study we carried out experiments in vitro and in vivo and investigated the effect of proline-rich polypeptide (PRP) on the proliferation and effectiveness of colony formation of MMSCs in vitro. Various routes and doses of PRP administration to rats increased the number of MMSCs in bone marrow and spleen. Our research revealed opposite effects of PRP on the proliferation of bone marrow stromal cells obtained from normal humans and stromal cells isolated from a human giant-cell tumour.

Comparative analysis of differentiation and behavior of human neural and mesenchymal stem cells In Vitro and In Vivo

Comparative analysis of differentiation of human neural and mesenchymal stem cells in tissue culture and after transplantation into the brain was carried out using the same antibody set. Neural stem cells differentiated into all types of neural cells, are retained after transplantation, migrate, and form reciprocal relationships with the recipient brain. Mesenchymal stem cells were incapable of neural development under conditions of common culturing or after transplantation and retained the fibroblast-like status. Recipient filaments grew into mesenchymal stem cell transplants containing no neural cells due to local changes in the extracellular matrix at the site of transplantation.

EFFECT OF CURETTAGE OF THE MEDULLARY CAVITY ON BONE MARROW STROMAL PRECURSOR CELLS

Changes in the number of stromal bone marrow precursor cells in guinea pigs after curettage of the medullary cavity were studied by cloning and monolayer cultures in vitro. Curettage was shown to remove about half of the fibroblast colony forming cells (FCFC) from the bone marrow. Later, the number of FCFC in the curetted limb fell to reach a minimum after 12 h. Starting from 24 h their number increased. By the 7th–12th day the number of FCFC reached the normal level, and by the 20th day it was 2.5 times higher than normal. The number of FCFC in the contralateral limb between 6 h and 20 days after curettage was 2–2.5 times greater than normal.

Effect of α-fetoprotein on the count of bone marrow and splenic stromal precursor cells and proliferation of their cultural descendants

The efficiency of cloning of stromal precursor cell increased more than 2-fold in splenic cultures and more than 3-fold in bone marrow cultures 24 h after injection of Profetal preparation to mice in vivo. The number of nucleated cells did not change in the bone marrow and slightly increased in the spleen. Addition of Profetal in vitro 2-fold decreased the efficiency of stromal precursor cells colony formation in mouse splenic cultures and dose-dependently decreased this process in bone marrow cultures derived from these animals, the maximum (5-fold) inhibitory effect was observed in a dose of 50 µg/ml. Addition of Profetal to cultured human bone marrow fibroblasts did not change the content of stromal fibroblasts in cultures. These data indicate the possibility of indirect effect of α-fetoprotein on the number of stromal precursor cell in hemopoietic and lymphoid organs.

Effects of Acoustic and EHF Impulses on Multipotent Stromal Cells during Formation of Bone Marrow Containing Heterotopic Organs in Tissue Engineered Constructions

We studied the effects of physical factors (acoustic impulses of laser-induced hydrodynamics, AILIH, and EHF-radiation) on the formation of heterotopic bone marrow organs. Suspension of precipitated mouse bone marrow cells was exposed to AILIH and EHF or their combinations (AILIH+EHF, EHF+AILIH). The developed tissue engineering constructions (gelatin sponges containing 107 nucleated bone marrow cells exposed to physical factors) were transplanted under the renal capsule of syngeneic mice. Analysis of newly formed hemopoietic organs was performed after 3 and 5 months. The total amount of hemopoietic cells, number of multipotent stromal cells, efficiency of colony formation from these cells, and weight of bone capsule of the transplants were measured. Microscopic study showed that 5-month transplants were significantly larger than 3-month transplants and contained 3-fold more hemopoietic cells (20-fold in the AILIH+EHF group). The number of multipotent stromal cells was maximum in EHF+AILIH group (by 2.2 times higher than in the control) and minimum in AILIH+EHF group. Exposure to EHF+AILIH had most pronounced effect on the formation of the bone marrow transplants. The weight of bone capsules more rapidly increased in gelatin sponges of 3-month transplants of EHF+AILIH and AILIH groups. These data suggest that the studied physical factors can be used for acceleration of rehabilitation process.

Comparison of the Efficiency of Transplantation of Bone Marrow Multipotent Mesenchymal Stromal Cells Cultured under Normoxic and Hypoxic Conditions and Their Conditioned Media on the Model of Acute Lung Injury

The therapeutic efficiency of intravenous injection of rat bone marrow multipotent mesenchymal stromal cells grown under conditions of normoxia and hypoxia (3% O2) and conditioned media from these cultures were compared on the rat model of acute lung injury induced by intraperitoneal injection of lipopolysaccharide. The best therapeutic efficiency was demonstrated by cells grown under hypoxic conditions. The effect of conditioned media was less pronounced and did not depend on the culturing conditions.