Functional Parameters of Physiological Systems of Laboratory Primates after Administration of Doxorubicin and Transplantation of Mesenchymal Stem Cells.

We studied physiological parameters of rhesus monkeys after administration of anthracycline antibiotic doxorubicin. Intravenous administration of the drug caused intoxication manifested in in an abrupt body weight loss, baldness, vomiting, and exicosis. Intoxication in monkeys determined by the damaging effects of doxorubicin on organs and tissues is also characterized by significant changes in the blood: leukopenia, thrombocytopenia, neutropenia, monocytopenia, lymphocytosis, and a sharp drop of CD20+ B cell content. The total protein and albumin content in the blood significantly decreased. A sharp increase in C-reactive protein was also accompanied by an increase in activity of proinflammatory cytokine IL-6. Transplantation of mesenchymal stem cells in some cases can significantly alleviate doxorubicin-induced damage to organs and maintain normal clinical status of monkeys after two injections of the drug. Late transplantation of stem cells does not have a protective effect and does not protect the animals from the damaging effects of doxorubicin. We found that the protective effect of mesenchymal stem cells depends on the dose of the drug, total number of cells, and the time of their transplantation. It should be noted that human and monkey mesenchymal stem cells produce similar regenerative effects, at least in the doxorubicin toxicity model.

Use of Mesenchymal Stem Cells for Possible Repair of Doxorubicin-Damaged Organs and Tissues in Experimental Monkeys.

Three injections of doxorubicin to rhesus macaques cause severe intoxication, characterized by anemia, cachexia, and degeneration of the viscera. The life span of monkeys injected with the drug and receiving after 24 h mesenchymal stem cell transplantation varied from 96 to 120 days in comparison with 50-74 days in animals receiving stem cells before doxorubicin. Controls received doxorubicin and saline; the lifespan of one monkey was 24 days, of the other - 1 year and 8 months. The increase in activity of proinflammatory cytokine IL-6 was paralleled by an increase in the level of C-reactive protein.

Allogenic cardiomyoblasts raised from human mesenchymal stem cells in the therapy of radiation cardiomyopathy and pericarditis: case report.

The use of triple systemic transplantation of cardiomyoblasts raised from the culture of allogenic bone marrow mesenchymal stem cells of a healthy donor according to the new medical technology licensed by Federal Service on Surveillance in Healthcare in the therapy of a patient with late radiation cardiomyopathy and radiation exudative pericarditis developed 45 years after radiation therapy for Hodgkin lymphoma. High efficiency of systemic transplantation of mesenchymal stem cells partially differentiated towards cardiomyocytes was demonstrated. The therapeutic effect persists for more than 2 years. Possible mechanisms of the therapeutic effect of this type of stem cells and the prospects of using cell therapy in the treatment of late radiation injuries of vital organs and tissues are discussed.

Possible aspects of using cultured stem cell of laboratory primates in experimental medicine.

We describe the methods of isolation and culturing of mesenchymal stem cells from 3 monkey species Macaca mulatta, Papio hamadryas, and Macaca fascicularis. Flow cytofluorometry showed that the cells do not express CD34, CD45, and HLA-DR, but most of them (78-98%) express CD90 marker. The cardioprotective effects of cultured mesenchymal stem cells in cardiomyopathy induced by administration of antitumor anthracycline drugs (doxorubicin).

Morphofunctional study of the therapeutic efficacy of human mesenchymal and neural stem cells in rats with diffuse brain injury.

We studied the effect of transplantation of human stem cells from various tissues on reparative processes in the brain of rats with closed craniocerebral injury. Combined treatment with standard drugs and systemic administration of xenogeneic stem cells had a neuroprotective effect. The morphology of neurons rapidly returned to normal after administration of fetal neural stem cells. Fetal mesenchymal stem cells produced a prolonged effect on proliferative activity of progenitor cells in the subventricular zone of neurogenesis. Adult mesenchymal stem cells had a strong effect on recovery of the vascular bed in ischemic regions.

Effects of (60)co whole-body gamma-irradiation in different doses on the distribution of (188)Re-labeled autologous mesenchymal stem cells in wistar rats after intravenous (systemic) transplantation during different periods after exposure.

The effects of whole-body gamma-irradiation in different doses on the distribution of (188)Re-labeled mesenchymal stem cells obtained by culturing of the rat bone marrow cells were studied in different organs and tissues of animals after intravenous (systemic) injection. Irradiation stimulated homing of (188)Re-labeled mesenchymal stem cells in organs and tissues of animals in comparison with this process in intact non-irradiated rats. The intensity of homing increased with increasing the irradiation dose and decreased with prolongation of the period between irradiation and systemic transplantation. It was hypothesized that increased migration of transplanted mesenchymal stem cells into irradiated organs and tissues can be caused by developing cell death processes. It seems that systemic transplantation of mesenchymal stem cells shortly after irradiation can be used for stimulation of reparative processes in damaged tissues.

[Systemic transplantation of autologous mesenchymal stem cells of the bone marrow in the treatment of patients with multidrug-resistant pulmonary tuberculosis].

The study undertaken 3 years ago examined the effect of systemic transplantation of autologous mesenchymal stem cells (MSC) in the complex therapy of 27 patients with pulmonary tuberculosis, including 15 patients with multidrug-resistant pulmonary tuberculosis and 12 with extensive drug resistance of Mycobacterium tuberculosis. All the patients were bacteria-discharging persons with disseminated destructive processes in lung tissue, most (n=17) of them had chronic fibrocavernous tuberculosis. In all the patients, previous long specific antituberculous treatment was ineffective or inadequately effective. After systemic MSC transplantation, 16 patients were followed up for 1.5-2 years or more and the remaining 11 patients for at least 6 months. After MSC administration, a positive clinical effect was observed in all 27 cases; bacterial discharge stopped in 20 patients after 3-4 months; resolution of sustained lung tissue cavities further occurred in 11 patients. At present, a persistent remission of a tuberculous process may be stated in 9 of the 16 patients in whom MSCs were transplanted 1.5-2 years, significant positive bacteriological and morphological changes are observed in 6 patients. Thus, inclusion of transplantation of the autologous MSCs propagated in the culture into a course of antituberculous therapy may be a promising procedure for enhancing the efficiency of therapy in patients with resistant forms of pulmonary tuberculosis.

Morphofunctional study of the therapeutic effect of autologous mesenchymal stem cells in experimental diffuse brain injury in rats.

Effects of systemic transplantation of mesenchymal stem cells obtained by culturing of autologous bone marrow on proliferative activity of cells and functional morphology of neurons after diffuse brain injury were studied in Wistar rats. Comparative analysis of the results indicated that systemic injection of mesenchymal stem cells in a syngeneic organism produced proliferotropic, angiogenic, and, presumably, neurotrophic effects. The therapeutic effect visually manifested on day 2 after intravenous injection of mesenchymal stem cells during the early period of reparative regeneration of ischemic cell and tissue structures of the brain. The neuroprotective effect of mesenchymal stem cells was more pronounced against the background of basic therapy.

Agar cultures of human clonogenic hemopoietic precursor cells for early diagnosis of some myeloproliferative diseases.

Growth characteristics of human hemopoietic cells in erythremia and chronic myeloid leukemia were studied using agar cultures with and without hemopoietic growth factors. Agar cultures, similarly to cultures on other semisolid media (plasma clot, methylcellulose) can be used for early differential diagnosis of polycythemia vera (erythremia) and secondary erythrocytosis: erythremia, but not erythrocytosis, is characterized by spontaneous (erythropoietin-independent) formation of colonies from erythrocyte precursor cells. Spontaneous colony formation from granulocyte-macrophage precursor cells can serve as an important test for early diagnosis of chronic myeloid leukemia. The study of colony formation from granulocyte-macrophage precursors and of the capacity of bone marrow cells to form colonies from hemopoietic stromal precursor cells revealed new characteristics of the studied myeloproliferative diseases. Presumably, spontaneous colony formation from erythrocytic and myeloid precursors should be regarded as a sign of tumor transformation of the studied hemopoietic cells.

[New developments in understanding the compensatory hematological reactions and their outcomes in local fractionated therapeutic gamma irradiation]. / Novoe v ponimanii kompensatornykh gematologicheskikh reaktsii i ikh iskhodov pri lokal'nom fraktsionirovannom terapevticheskom gamma-obluchenii.

We have carried out a study of the bone marrow status in both irradiated and non-irradiated zones of 56 patients with stage I-II Hodgkin's disease in complete 9-12 (33 patients, group 1) and 18-23 (23 patients, group 2) year remission after therapeutic irradiation of the supradiaphragmatic lymphatic collectors at a dose of 40 Gy with irradiation of the spleen (33 patients) or splenectomy (23 patients). The total count of myelokaryocytes, myelogram, a relative and absolute content of lymphoid cells, immature granulocytes and elements of erythroid series were calculated in the aspirates from the exposed to radiotherapy sternum and non-irradiated upper portion of the ileum. The number of granulocyte-macrophage (CFU-GM) and stromal (CFU-F) precursor cells were defined using in vitro culture technique. There was a complete annihilation of the bone marrow in the irradiated zones, when the dose exceeded 35 Gy in 3-4 weeks. The concentration of myelokaryocytes, immature granulocytes, erythronormoblasts, CFU-GM, CFU-F in non-exposed bone marrow were significantly lower in all patients of group 2 than in normal subjects and in group 1 patients. Absolute lymphoid count in patients with 18-23 year remission was found to be normal but was considerably reduced in comparison to patients of group 1. These changes may be the result of the previous hyperactivity of the non-irradiated bone marrow which could be a cause of stem cell compartment depletion. The differential calculation of compact and diffuse subpopulations of CFU-F revealed a significant reduction of compact colony-forming CFU-F in both irradiated and unexposed bone marrow. Almost all the stromal precursor cells from irradiated zone formed diffuse colonies in cultures. These results confirm experimental data concerning greater radiosensitivity and proliferative potential of CFU-F, forming compact colonies versus diffuse colony-forming CFU-F. Aplasia of the irradiated bone marrow and hypoplasia of the non-irradiated bone marrow 18-23 years after radiotherapy completion coexisted with normal circulating CFU-GM and granulocyte blood count suggesting a compensatory mechanism involving a mitotic amplification between the progenitor cell and the final differentiated cell.

[The effect of the gas hypoxic mixture GHM-8 on the capacity of the stromal clonogenic cells (CFU-F) in rat bone marrow for postradiation recovery]. / Vliianie gazovoi gipoksicheskoi smesi GGS-8 na sposobnost' stromal'nykh klonogennykh kletok (KOE-F) kostnogo mozga krys k postradiatsionnomu vosstanovleniiu.

The effect of gas hypoxic mixture, containing 8% of O2 (GHM-8), on the ability of cell precursors of haemopoietic stroma (which form colonies (clones) of fibroblasts (CFU-F) in a culture, and are present in the bone marrow of adult rats) to repair potentially lethal and sublethal radiation damages has been investigated. The recovery of CFU-F from potentially lethal damages, that was studied after their delayed survival in a culture following irradiation of animals, proceeds at nearly the same rate in cells irradiated both in the air and in hypoxic conditions (GHM-8). Fractionated irradiation reduces the radioprotective effect of GHM-8 for CFU-F, particularly for the radioresistant subpopulation; the ability of CFU-F to recover from sublethal radiation damages decreases.

[The capacity of rat clonogenic precursor cells of the hemopoietic stroma (CFU-F) to recover from radiation damages]. / Sposobnost' krysinykh klonogennykh kletok-predshestvennikov gemopoéticheskoi stromy (KOE-F) k vosstanovleniiu ot radiatsionnykh povrezhdenii.

The ability of cell precursors of the haemopoietic stroma (CFU-F), that are present in the bone marrow of adult rats, to recover from potentially lethal and sublethal radiation damages has been investigated. The highest reparability, with respect to potentially lethal damages, is displayed by the most radioresistant CFU-F population, that forms loose colonies (clones) in a culture; the slope of the dose-response curve, not the extrapolation number, changes, and heterogeneity of the CFU-F population is observed. The results obtained confirm the presence of heterogeneity in the population of CFU-F, that was revealed in studying their radiosensitivity by the formation of dense and loose fibroblast colonies in a culture.

[Radiosensitivity of cells-precursors of hemopoietic stroma (CFU-F) in the rat bone marrow under the effect of 60Co gamma irradiation in various conditions]. / Radiochuvstvitel'nost' kletok-predshestvennikov gemopoéticheskoi stromy (KOE-F), soderzhashchikhsia v kostnom mozge krys, pri deistvii gama-izlucheniia 60Co v razlichnykh usloviiakh.

Conditions have been developed for cloning cells-precursors of rat bone marrow haemopoietic stroma, that form in culture dense and sparse fibroblast colonies (CFU-F) at a plating efficiency of 10(-4). Radiosensitivity of rat bone marrow CFU-F, with 60Co-gamma-irradiation in vitro, is characterized by the values of Do and n of 1.87 Gy and 1.4 respectively for all clones; 0.65 Gy and 6.7 for dense clones, and 4.27 Gy and 1.0 for sparse clones. This confirms the observed heterogeneity of CFU-F population consisting of highly radiosensitive and radioresistant subpopulations. The parameters of rat bone marrow CFU-F are nearly the same with irradiation both in vivo and in vitro; with in situ irradiation, the oxygen effect comes into play in a radiosensitive subpopulation of CFU-F; the OER values are 1.6, 2.6 and 0.9 for all, dense and sparse clones respectively.

[The radioprotective effect of hypoxia on clonogenic cells of rat bone marrow stroma (KOE-F)]. / Radiozashchitnoe deistvie gipoksii na klonogennye kletki stromy kostnogo mozga (KOE-F) krys.

A comparative study was made on the survival rate of cell-precursors of haemopoietic stroma, that form, in a rat bone marrow culture, colonies (clones) of fibroblasts (CFU-F) after gamma-irradiation of animals in the air or in a gas hypoxic mixture, containing 8% of O2 (GHM-8). Irradiation in GHM-8 was shown to increase the survival rate of CFU-F by 1.7 times (as compared to exposure in the air) as estimated by the total number of colonies that are formed in a culture; the radioprotective effect of GHM-8 was more pronounced for CFU-F which form dense colonies: DMF for dense and loose clones was 2.4 and 1.6 respectively.

[Thermosensitivity of clonogenic cells and the induction of thermal tolerance]. / Termochuvstvitel'nost' klonogennykh kletok i induktsiia v nikh termotolerantnosti.

Clonogenic cells of some normal tissues (CFU-GM of the bone marrow of mice, CFU-F of the bone marrow of rats, CFU-GM and CFU-F of the bone marrow of dormice) as well as tumor clonogenic cells of the ascitic variant of ascitic Ehrlich carcinoma were characterized by similar thermosensitivity during their heating under in vitro conditions whereas thermosensitivity in clonogenic cells of solid tumors (LLC, solid Ehrlich ascitic carcinoma) turned out twice as high. A possibility of inducing thermotolerance in these cells by various modes was shown.