Isolation and characterization of mesenchymal stem cell population entrapped in bone marrow collection sets.

Bone marrow is an important source of mesenchymal stem cells (MSCs), and a promising tool for cytotherapy. MSC utilization is limited by low cell yields obtained under standard isolation protocols. Herein, used bone marrow collection sets were evaluated as a valuable source of MSCs.

[Intensive therapy with paclitaxel (Taxol) and cyclophosphamide followed by administration of G-CSF as a mobilization regimen in patients with breast carcinoma and indications for autologous hematopoietic cell transplantation].

Cyclophosphamide (4 g/m2) and paclitaxel (Taxol) (175, 200 or 250 mg/m2) therapy with subsequent administration of G-CSF (10 micrograms/kg) has been used as intensification and as mobilization therapy for patients with breast cancer. This regimen was used in 19 patients, as part of adjuvant therapy in 14 and as part of therapy of metastatic disease in five. Median number of collected CD34+ cells was 17.

[Manipulation of hematopoietic stem cell grafts and their use in clinical practice].

Autologous stem cell transplantation has been successfully used in treatment of various hematological malignancies and solid tumors in children and adults. Published data have confirmed that bone marrow harvests and peripheral blood stem cell collections frequently contain a significant number of tumor cells. Contaminating tumor cells can contribute to the disease relapse in posttransplant period, so attempts are made to eliminate contaminating tumor cells from autografts.

Comparison of two different methods for CD34+ selection and T cell depletion in peripheral blood stem cell grafts--our experiences with CellPro, E rosetting and CliniMACS technique.

The aim of this study was to establish a suitable method for in vitro T cell depletion in peripheral blood stem cell grafts for mismatched/haploidentical transplantation in children and adults with severe hematological disorders and for autologous transplantation in patients with autoimmune diseases refractory to conventional immunosuppressive treatment. Two different selection techniques have been used: CD34+ selection using immunoaffinity columns (CellPro Ceprate) followed by T cell depletion by E-rosetting or CD34+ selection using submicroscopic paramagnetic beads (CliniMACS device) with T cell depletion in a one step procedure. The mean purity and recovery of CD34+ cells and efficiency of T cell removal in the final product were compared.

The efficiency of PBPC collections and the relationship to the precollection concentration of CD 34+ cells in blood.

Transplantations of peripheral blood progenitor cells (PBPC) are able to assure a complete haematopoietic and immunologic reconstitution. The efficient mobilization of progenitor cells into peripheral blood is the main factor responsible for quality of the graft as well as timing and technique of collections. The aim of the present paper was to find the optimum time for starting PBPC collections and consequently to minimize the number of procedures required.

[Separation of hematopoietic progenitor cells from peripheral blood in patients with hemato-oncologic malignancy].

Background: Transplantations of haematopoietic progenitor cells from peripheral blood (PBPC) are able to ensure haematopoietic and immunological reconstitution as well as stable long term engraftment. Autologous PBPC are administered after previous myeloablative chemotherapy to patients with haematological and non-haematological malignancies. The objective of the submitted study was to follow-up the results of autologous separations of PBPC in patients with a good effect of mobilisation therapy.

Transplantation of allogeneic peripheral blood progenitor cells--a single centre experience.

13 patients have been transplanted at Institute of Hematology and Blood Transfusion since 1995 using allogeneic PBPC either alone or with bone marrow as a source of progenitor cells. All donors were G-CSF mobilised HLA identical family members. PBPC harvests were performed on D 4,5, (6) of G-CSF administration.

[Allogenic transplantation of hematopoietic stem cells in the treatment of children with high-risk acute leukemia].

Background: Most children with acute lymphoblastic leukemia (ALL) and increasing number of children with acute myelogenous leukemia (AML) are currently cured with conventional chemotherapy. Despite of this success there is a subset of patients with high-risk features at diagnosis who are predisposed to a very high risk of relapse. Relapse of AML and early bone marrow relapse of ALL can not be cured by conventional chemotherapy.

[Purging of hemopoietic progenitor cells in autologous transplantation].

The authors describe the results of purification of bone marrow and peripheral progenitor cells (PBPC) for clinical transplantations. Vepeside was used to purify in 1990-1995 a total of 41 bone marrows of adults and children. Of these 23 were transplanted.

[Allogenic transplantation of bone marrow in childhood].

Background: Bone marrow transplantation has become the therapeutic method in some forms of malignant haemotopoietic diseases, malignant tumours, inborn errors of metabolism and immunodeficiency states. The objective of the presented work is the analysis of 40 allogenic bone marrow transplantations in children made in 1989-1994.

Methods And Results: Bone marrow transplantation was made in 40 children (26 boys and 14 girls), mean age 10.

[Results of bone marrow transplantation at the Institute of Hematology and Blood Transfusion in Prague].

The first allogenic bone marrow transplantation (TKD), when for the preparation whole body irradiation was used, was implemented in the Institute of Haematology and Blood Transfusion (UHKT) in Prague in 1986. Before June 1992 36 TKD were performed incl. 28 allogenic, 2 syngenic and 6 autologous.

Induction of NK and LAK activities in human lymphocyte culture by a cytosol fraction from leukemic myeloblasts and by monoclonal antibody CD 3.

The stimulating effect of cytosol fraction (F3) isolated from human myeloblasts (m.w. ranging from 30 to 100 kDa) and monoclonal antibody CD 3 (MEM 57) was tested on NK and LAK cell activities in peripheral mononuclear cells (PMNC) of normal donors and leukemic patients.

Positive effect of direct current on cytotoxicity of human lymphocytes.

Normal or leukemic human lymphocytes treated with direct current (DC) showed enhanced antileukemic cytotoxicity. The enhancing effect of DC-treated lymphocytes was dependent on current density and time exposure. A desirable effect was achieved with current densities ranging from 5 to 10 mA/cm2 at a short exposition time (5--10s).

Human leukocyte markers defined by monoclonal antibodies. I. Expression of X-hapten structure on cells of myeloid lineage.

We report on the characterization of four monoclonal antibodies which were prepared against membrane markers of human myeloid lineage. Fusion, isolation of hybridoma cells and their cloning and testing of the monoclonal antibodies by indirect immunofluorescence and FACS 440 analysis were performed by means of standard procedures. The results indicate that the monoclonal antibodies have a specificity against membrane markers of human myeloid lineage (exactly promyelo-granulocytes).

Effect of a synthetic poly N-(2-hydroxypropyl)methacrylamide (Duxon) on haemopoiesis and graft-versus-host reaction.

A synthetic polymer, Duxon, was developed and tested as a substitute of blood plasma for transfusion purposes. Tests of this preparation included a test for its influence on the haemopoietic stem cells and the graft-versus-host reaction (GVHR). A single dose or repeated doses of Duxon did not reduce the number of pluripotent haemopoietic colonies (CFU-S) in mice and short-term incubation of cells from haemopoietic organs of mice with Duxon resulted in a slight, yet significant, increase in the number of CFU-S.

Influence of the incubation of cells with zinc and lithium ions on GVH reactivity of cells and on their ability to form haemopoietic colonies.

Mouse spleen and bone marrow cells were incubated for 2 h with ZnCl2 and Li2SO4 at different concentrations and tested for the ability to evoke the graft-versus-host reactions (GVHR) and to form pluripotent haemopoietic colonies. ZnCl2 at concentrations 5 X 10(-6) to 5 X 10(-4) M inhibited the regional GVHR. At a concentration of 5 X 10(-6) ZnCl2 also inhibited the ability to elicit the systemic GVHR in irradiated mice.

Derivatives of benzo(c)fluorene. XI. Effect of Benfluron on hemopoietic stem cells.

The effect of Benfluron-- 5-(2-N,N-dimethylamino-ethoxy)-7-oxo-7H-benzo(c)fluorene hydrochloride--on hemopoietic stem cells was tested by the production of spleen cell colonies (CFU-S) in irradiated mice following application of bone marrow cells and by the production of hemopoietic colonies (CFU-C) in semisolid agar. The reduced numbers of CFU-S were found in mice applied Benfluron or Benfluron-treated bone marrow cells. The increased numbers of CFU-S, however, were found in mice receiving bone marrow cells from Benfluron-treated donors.

Derivatives of benzo(c)fluorene. X. Inhibitory effect of Benfluron on cellular immunity.

The effect of a cytostatic drug Benfluron-- 5(2-N,N-dimethyl-amino-ethoxy)-7-oxo-7H-benzo(c)fluorene hydrochloride was tested in mice on skin graft survival, graft-versus-host reaction (GVHR) and for mitogenic stimulation of human lymphocytes. Application of Benfluron resulted in a prolonged skin graft survival. The regional and systemic GVHR was potentially inhibited by p.

The influence of methylpropionic acid and pyridazinone-3 derivatives on some immunologic and hemopoietic functions.

There was studied the influence on the cell-mediated and humoral response in vivo manifested by selected methylpropionic acid and pyridazinone-3 derivatives which had been found to possess strong immunotropic effects in the in vitro screening previously. It was shown that the compounds were generally poorly tolerated by animals, and they exerted only weak suppressive effects on antibody production, the contact hypersensitivity and survival of skin grafts. This immunosuppressive activity was accompanied by a slight decrease in the number of spleen colony forming cells (CFU-s).

Immunosuppressive effects of bovine seminal fluid fractions with ribonuclease activity.

Using different isolation procedures (after acidification and saturation with 3 M ammonium sulphate) three fractions were isolated from bull seminal vesicle fluid and assayed for their effects on cell immunity in vitro and in vivo. Two of these preparations (ZS RNase and AS RNase) possessing a high level of ribonuclease activity at concentrations of 50 micrograms/ml showed inhibitory effects (up to 80%) on 3H-thymidine incorporation into the DNA of mitogen-or antigen-stimulated human lymphocytes. The third preparation (3M-P) possessing low ribonuclease activity showed lesser inhibitory effects.

Inhibitory effects of 5-azapyrimidine nucleosides on cellular immunity.

The effect of 5-azacytidine (5-AzCR) and 5-aza-deoxycytidine (5-AzCdR) on the survival of skin grafts in mice and rats, the action of these drugs on regional GVH reaction, as well as the formation of haemopoietic colonies (CFU-5) in the spleen were studied. Both drugs prolonged the life span of skin grafts when administered 24 hr before transplantation, or on the 4th post-transplantation day. However, they were little effective when injected 24 hr after skin grafting, or after induction of the regional GVHR.

The effect of Damvar on the formation of hemopoietic splenic colonies in mice, and on the peripheral blood picture in rats and dogs.

The present paper deals with the effect of Damvar on hemopoiesis. The test of formation of hemopoietic splenic colonies (CFU-S) was used in mice, the peripheral blood picture and bone marrow differential after long-term administration of Damvar was tested in rats and dogs. The bone marrow cells from mice after a single dose of Damvar or cells incubated with Damvar in vitro showed no reduction of colony-forming activity.

Effect of the treatment of parental cells in vitro with anti-T antibody on bone marrow transplantation in F1 hybrid mice.

Anti-T antibody was obtained from xenogeneic antithymocyte serum by adsorption and elution with brain immunosorbent. Bone marrow and spleen cells from the parental A strain were treated in vitro with anti-T antibody and complement or with the original ATS and complement. So treated cells were assayed for the ability to induce a local and a total GVH reaction in (A X C57BL)F1 hybrid mice, to prolong the survival of lethally irradiated, identical F1 hybrid mice and to form haemopoietic colonies in spleens of the syngeneic irradiated recipients.