Growth and differentiation of bone marrow hematopoietic cells in mice bearing Ehrlich ascite tumor and treated with Dicyclopentadienildichiorotitanium (IV)

In this work we have:investigated the growth and differentiation of bone marrow stem cells in mice bearing Ehrlich ascites tumor-and treated with three dose-regimens of Dicyclopentadienyldichlorotitanium (IV) (DDCT). We also: studied the presence of colony stimulating factors In the serum of PDCT-treated animals as well-as the effects-of the drug on the survival of the tumor-bearing mice. The-results demonstrated that the myelosuppression developed in the tumor-bearing animals is prevented by the administration:of 1, 2 or 3 doses of 15 mg/kg DDCT. In the treatment with three doses, however, 23 % of the animals died. Moreover, DDCT treatment in normal animals resulted in increased numbers of CFU-GM. We observed the presence of stimulating factors in the serum of drug-treated animals which induced the growth and differentiation of bone marrow progenitor cells from normal animals in vitro. on the other hand, in vitro addition of the drug to these cultures had no effect. Thus, we conclude that the drug protects against the myelosuppression induced by the tumor and that this protection may be related to an indirect action of the drug. (C) 1998 International Society for Immunopharmacology. Published by Elsevier B.V. Ltd.

Progesterone upregulates GATA-1 on erythroid progenitors cells in liquid culture

Steroids hormones modify the hematological features of homozygous sickle cell disease, including the levels of fetal hemoglobin. We used semi-quantitative RT-PCR analysis of GATA-1, GATA-2, NF-E2, and gamma-globin mRNA levels in a two-phase liquid culture system of human adult erythroid cells in order to assay the effect of progesterone upon gene expression. The levels of expression of GATA-1 and gamma-globin mRNA were significantly increased in cells treated with progesterone compared to untreated cells (1.7- to 2.0-fold). Progesterone treatment did not produce any stimulatory effect upon GATA-2 and NF-E2 mRNA expression. Differences in the synthesis of HbF protein could not be detected by flow cytometry, although we observed a small difference in mean intensity fluorescence between cells treated and cells untreated with progesterone on days 7 and 9. Using anti-transferrin receptor and anti-glycophorin A antibodies, we verified that addition of progesterone did not cause any change in erythroid proliferation and differentiation. In conclusion, it is possible that the increased expression of gamma-globin mRNA after progesterone treatment observed in this study may be related to the increased GATA-1 mRNA expression. Interactions of the steroid receptors with the basal transcriptional machinery and with transcription factors might mediate their transcriptional effects. (C) 2002 Elsevier B.V. (USA).

The effect of CpG-ODN on antigen presenting cells of the foal

Background: Cytosine-phosphate-guanosine oligodeoxynucleotide (CpG-ODN) has been used successfully to induce immune responses against viral and intracellular organisms in mammals. The main objective of this study was to test the effect of CpG-ODN on antigen presenting cells of young foals. Methods: Peripheral blood monocytes of foals (n = 7) were isolated in the first day of life and monthly thereafter up to 3 months of life. Adult horse (n = 7) monocytes were isolated and tested once for comparison. Isolated monocytes were stimulated with IL-4 and GM-CSF (to obtain dendritic cells, DC) or not stimulated (to obtain macrophages). Macrophages and DCs were stimulated for 14-16 hours with either CpG-ODN, LPS or not stimulated. The stimulated and non-stimulated cells were tested for cell surface markers (CD86 and MHC class II) using flow cytometry, mRNA expression of cytokines (IL-12, IFNα, IL-10) and TLR-9 using real time quantitative RT-PCR, and for the activation of the transcription factor NF-κB p65 using a chemiluminescence assay. Results: The median fluorescence of the MHC class II molecule in non-stimulated foal macrophages and DCs at birth were 12.5 times and 11.2 times inferior, respectively, than adult horse cells (p = 0.009). That difference subsided at 3 months of life (p = 0.3). The expression of the CD86 co-stimulatory molecule was comparable in adult horse and foal macrophages and DCs, independent of treatment. CpG-ODN stimulation induced IL-12p40 (53 times) and IFNα (23 times) mRNA expression in CpG-ODN-treated adult horse DCs (p = 0.078), but not macrophages, in comparison to non-stimulated cells. In contrast, foal APCs did not respond to CpG-ODN stimulation with increased cytokine mRNA expression up to 3 months of age. TLR-9 mRNA expression and NF-kB activation (NF-kB p65) in foal DCs and macrophages were comparable (p > 0.05) to adult horse cells. Conclusion: CpG-ODN treatment did not induce specific maturation and cytokine expression in foal macrophages and DCs. Nevertheless, adult horse DCs, but not macrophages, increased their expression of IL-12 and IFNα cytokines upon CpG-ODN stimulation. Importantly, foals presented an age-dependent limitation in the expression of MHC class II in macrophages and DCs, independent of treatment. © 2007 Flaminio et al; licensee BioMed Central Ltd.

Influence of endogenous and synthetic female sex hormones on human blood cells in vitro studied with comet assay

The comet assay has been conducted with numerous cell lines to assess in vitro genotoxicity. In order to use the comet assay as part of an in vitro test for evaluating genotoxicity, however, there are cell-specific factors that need to be better understood. In this present study we have evaluated some factors that may impact upon the DNA damage detected in whole blood (WB) cells and lymphocytes (ILs). Experiments were conducted comparing responses of both cells, and investigating the effects of the female hormonal cycle, and oral contraceptive (OC) use on DNA damage detection in the in vitro comet assay, at three sampling time. No significant differences were detected in the basal levels of DNA damage detected in ILs and WB cells from women OC users and non-users and from men. Basal DNA damage in ILs was unaffected by gender and stage of the menstrual cycle or the stage of the treatment schedule. Our results also indicated that the H2O2 induces DNA damage in human lymphocytes independently of gender, low-dose OC use and hormonal fluctuation. However, data showed that in 3rd sampling of menstrual cycle, lymphocytes were more resistant to H2O2-induced DNA damage than those from OC users and men. © 2007 Elsevier Ltd. All rights reserved.

Cell Therapy with Bone Marrow Mononuclear Cells in Elastase-Induced Pulmonary Emphysema

Emphysema is characterized by destruction of alveolar walls with loss of gas exchange surface and consequent progressive dyspnea. This study aimed to evaluate the efficiency of cell therapy with bone marrow mononuclear cells (BMMC) in an animal model of elastase-induced pulmonary emphysema. Emphysema was induced in C57Bl/J6 female mice by intranasal instillation of elastase. After 21 days, the mice received bone marrow mononuclear cells from EGFP male mice with C57Bl/J6 background. The groups were assessed by comparison and statistically significant differences (p & 0. 05) were observed among the groups treated with BMMC and evaluated after 7, 14 and 21 days. Analysis of the mean linear intercept (Lm) values for the different groups allowed to observe that the group treated with BMMC and evaluated after 21 days showed the most significant result. The group that received no treatment showed a statistically significant difference when compared to other groups, except the group treated and evaluated after 21 days, evidencing the efficacy of cell therapy with BMMC in pulmonary emphysema. © 2012 Springer Science+Business Media New York.

Produção de embriões por fecundação in vitro e ativação partenogenética visando o isolamento de células tronco embrionárias felinas

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Imunofenotipagem de subpopulações de linfócitos-T no sangue do cordão umbilical de eqüinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Padronização da coleta e quantificação de células-tronco hematopoéticas no sangue do cordão umbilical de cães

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Resposta tecidual de dentes reimplantados utilizando plasma pobre em plaquetas, plasma rico em plaquetas e células-tronco da medula óssea: estudo em cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Isolamento, caracterização e diferenciação de células tronco embrionárias e mesenquimais de equinos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Do progenitor cells from different tissue have the same phenotype?

The purpose of this work was to validate isolation methods for sheep mesenchymal stem cells (MSC) from different sources and to explore the hypothesis that MSC exhibit markers of the same phenotype independent from tissue source. Cells derived from ovine bone marrow, synovial membrane and adipose tissue were characterized using the following markers: CD44, CD45, CD11b and MHC-I. The isolated MSC were cultivated, went through osteogenic, chondrogenic and adipogenic differentiation, and were characterized by flow cytometry using mouse anti-ovine CD44, CD45 and MHC-I monoclonal antibody (mAb), and mouse anti-bovine CD11b mAb. Ovine MSC from all three sources differentiated under chondorgenic, osteogenic and adipogenic conditions. Also, MSC from the three tissues were found to express CD44 and MHC-I but lack of CD11b and CD45. The results obtained revealed that our isolation methods for the different tissues tested are valid and that MSC from the three sources studied have same immunophenotic characteristics. (C) 2014 Published by Elsevier Ltd. All rights reserved.

Immunodetection of cells with a CD44+/CD24-phenotype in canine mammary neoplasms

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Novel Chemically Modified Bacterial Cellulose Nanocomposite as Potential Biomaterial for Stem Cell Therapy Applications

Bacterial cellulose (BC) has become established as a remarkably versatile biomaterial and can be used in a wide variety of applied scientific applications, especially for medical devices. In this work, the bacterial cellulose fermentation process is modified by the addition of hyaluronic acid and gelatin (1% w/w) to the culture medium before the bacteria is inoculated. Hyaluronic acid and gelatin influence in bacterial cellulose was analyzed using Transmission Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). Adhesion and viability studies with human dental pulp stem cells using natural bacterial cellulose/hyaluronic acid as scaffolds for regenerative medicine are presented for the first time in this work. MTT viability assays show higher cell adhesion in bacterial cellulose/gelatin and bacterial cellulose/ hyaluronic acid scaffolds over time with differences due to fiber agglomeration in bacterial cellulose/gelatin. Confocal microscopy images showed that the cell were adhered and well distributed within the fibers in both types of scaffolds.

Caracterização, criopreservação e diferenciação in vitro de células tronco, derivadas de amniótica e gelatina de Wharton canina, visando a formação de um banco de células tronco

Pós-graduação em Biotecnologia Animal - FMVZ