Laboratory for Research Teaching (LRT)

The LRT is a new lab of 36,8 m2 equipped with research infrastructure in molecular and cellular biology that is been used for the development of introductory research projects by basic schools teachers and students.

 

 

 

 

Cellular Culture

The Cellular Culture Lab. performs researches on mesenchymal stem cells, hematopoietic and endothelial progenitor cells. These basic studies of biological characterization of mesenchymal stem cells contribute to the investigation of their clinical use in regenerative medicine and tissue engineering.

The Laboratory also performs researches to verify the expansion capacity of in vitro hematopoietic progenitor cells and the differentiation potential in dendritic cells.

The evaluation of the vasculogenic potential of endothelial cells EPC is also investigated. These preclinical studies collaborate with the characterization concerning the revascularization potential of an injured organ after vascular trauma such as ischemia or myocardial infarction.

 

Cellular Biology

The Laboratory conducts research on somatic stem cells, also known as adult stem cells, such as mesenchymal stem cells, hematopoietic and endothelial progenitor cells. These studies comprise the isolation, culture, expansion and the functional and immunophenotypical characterization of these cells, contributing to a better understanding of the potential clinical usage. The Laboratory also performs assay of spontaneous formation of erythroid colonies to help the Polycythemia Vera diagnosis.

 

 

 

Cryobiology Laboratory

The main function is to cryopreserve the cells for transplantation. The products which contain hematopoietic stem cell (HSC) such as bone marrow, umbilical cord blood and mobilized peripheral blood are processed to keep their biological function until the moment of use, what may happen years after the collection. The cell products are mixed with a cryoprotective solution, and then, the cell suspension is exposed progressively to lower temperatures until it reaches -80 to -100 ºC, then it is transferred to a container with liquid nitrogen (storage at -160 ºC) or to mechanic freezers (-80 to -150 ºC).
In addition to the cryopreservation, this Laboratory processes bone marrow with the purpose to reduce the volume of plasma and/or red blood cells in a context of ABO incompatibility.

 

Cytogenetics and Immunofluorescence

The Laboratory works on two main fields: 1) Molecular and Classical Cytogenetics (FISH) over oncohematological diseases (CML and multiple myeloma) and solid tumors (gliomas), and 2) the investigation of the cytogenetical  behavior of stem cells of adults expanded in vitro and their behavior in vivo.

 

 

 

 

Flow Cytometry

Through the flow cytometry technique, the Laboratory performs testing such as quantification lymphocyte subpopulations of HIV patients, progenitor stem cells quantification for allogeneic or autologous transplant, platelet syndromes determination, peripheral blood subpopulation quantification, and other methodologies. This technique allows the measurement of the chemical and physical characteristics in particles or in cells in suspension.

The Laboratory also supports researches performed on mesenchymal, endothelial, and dendritic cells. The Laboratory has the Flow Cytometer, which is able to perform the “cell sorter” (separation of subpopulation), to perform simultaneous analyses on seven different fluorescences.

 

Gene Transfer

The Gene Transfer Laboratory conducts researches on two main fields and a third one on embryonic stem cells has been recently initiated. The first field is the development of a bioprocess for the production of recombinant coagulation proteins for the treatment of patients with hemophilia A or B.
At present, the main challenges are the large-scale production of these proteins through bioreactor techniques and the development of animal models for hemophilia using cell-based therapy.
The second field of the research is the evaluation of therapeutic properties of mesenchymal stem cells (MSC) and pericytes, in particular the angiogenic characteristics and immunologic potentials. Both properties are evaluated by real time PCR and by the development of in vivo assay using NOD-scid mice for human stem cell engraftment. Also, the Institution developed two mouse tumor models to investigate the role of MSC in the tumor microenvironment. In this second field, the current challenge is the characterization of more primitive MSC to understand the molecules and signaling pathways involved in vascular lineage commitment which will contribute to design effective stem cell-based therapies.

 

Molecular Genetics and Bioinformatics

The major focus of our group is to understand the molecular mechanisms involved in genetic diseases and cancer development. We are using whole genomic analysis to identify and characterize genetic program/gene pathways that drive tumorigenic processes, and biomarkers of diagnosis, prognosis and therapeutic orientation. We are also interested in the characterization of biological properties of cancer stem cells (CSCs), which present self-renewal capacity, unlimited proliferative potential, differentiation competence, and are extremely resistant to radio and chemotherapies. For this purpose, we are applying next generation sequencing in order to profile gene expression, characterize epigenetic alterations, detect chromosomal rearrangements, and single nucleotide variations in coding and regulatory gene regions (RNAseq, ChIP-Seq, DNA methylation, Exome, and Paired-End protocols) in several solid tumors including: head and neck carcinoma, colorectal carcinoma, glioblastoma multiforme and osteosarcoma.

 

 

 

 

 

 

Molecular Biology

The Molecular Biology Laboratory performs researches on the diagnosis and molecular characterization for human retroviruses such as HIV, HTLV and hepatitis. During the last years, it has improved the investigation area, including researches related to stem cells properties and their modifications, erythrocyte antigens genotyping and gene polymorphism analyses.

The Laboratory, through Molecular Biology techniques, has the purpose of producing and developing tools for the diagnosis and therapy in areas such as Hemotherapy and Cell Therapy (molecular diagnosis of human retroviral infections, blood group genotyping).

 

Protein Chemistry Center

The Protein Chemistry Center develops several studies on function-structure of proteins in experimental model of cancer (using cell lineages and small animals) and human stem cells. The aim of these studies is to offer means to clear up the role of macromolecules (proteins) either in physiologic and pathologic processes. The goal of the research is the discovery of new biomarkers of diseases like cancer for early diagnosis and new targets for therapy development.
In stem cells, the Center develops research to understand the cell differentiation in different types of tissues which will be used in the future for regenerative medicine. These studies have been developed by using techniques such as dimensional electrophoresis, capillary liquid chromatography and mass spectrometry.
In addition, the Center develops an important service for the community in the amino acid analysis offering support to the General Hospital (FMRP-USP) and Associação de Pais e Amigos dos Excepcionais (APAE) for the diagnosis of inborn metabolism errors (phenylketonury, tyrosinemy, etc.).

 

Umbilical Cord Bank

The umbilical cord blood (UCB), for being rich in hematopoietic stem cells, may be an alternative source for transplants, instead of bone marrow, mainly for those patients without donors in the family.
For this reason, a public bank network of umbilical cord blood, named BrasilCord, has been implemented in Brazil, providing medical care for the Brazilian. It consists of 12 banks spread in the five regions of the country.
At present, banks from Instituto Nacional do Câncer (INCA), Hospital Israelita Albert Einstein, Universidade de Campinas (UNICAMP), and Regional Blood Center of the General Hospital of the Medical School of Ribeirão Preto (CRH – HCFMRP/USP) have taken part in this network. The last three ones, through a partnership, established the RedeCord, which is a public network of the UCBB from the state of São Paulo.
The umbilical cord blood is collected after the childbirth, stored in special bags, evaluated through various patterns, such as the amount of cell, contamination by infectious agents, and so on. Once the analysis has been done, the units of UCB in accordance with the current rules are processed and stored in a container with liquid nitrogen.

 

 HLA Genotyping Lab


The Human Leukocyte Antigen (HLA) Laboratory provides histocompatibility testing for kidney and bone marrow transplants, working 24h a day. Donor-recipient pairs are analyzed for HLA antigens compatibility. The HLA Laboratory performs compatibility testing for: 1) all cadaver donors of São Paulo State; 2) living kidney and bone marrow donors (family members) for transplantation in the University Hospital of Medical School of Ribeirão Preto, University of São Paulo; 3) volunteer donor banks of bone marrow and umbilical cord blood, REDOME and Brasilcord respectively, which are centralized at the Instituto Nacional do Câncer of Rio de Janeiro.

 

Hematology

Our laboratory is interested in the study of human telomere diseases. We are trying to understand the molecular and cellular mechanisms through which short and dysfunctional telomeres lead to adult stem cell failure, fibrotic diseases, and cancer. For this purpose, we use telomerase-deficient murine models and induced pluripotent stem (iPS) cells to model disease and to identify abnormal pathways derived from deficient telomere repair. We also screen patient samples for telomere length and mutations in novel genes that may be associated with telomere biology.

 

Molecular Morphophysiology and Development

The Molecular Morphophysiology and Development Laboratory (LMMD), located in Pirassununga (100 km from Ribeirão Preto), is part of the Basic Sciences Department at the Faculty of Animal Science and Food Engineering (FZEA) – University of São Paulo (USP). Established on April/2001 with the sponsor and leadership of Prof. Dr. Flávio V. Meirelles, the LMMD currently congregates 9 associated researchers (professors and post-doc fellowships) forming a team-based collaborative research group. The collective aims of the research developed at the LMMD include the understanding of mechanisms of embryo development and pluripotent cell biology, nuclear reprogramming and cytoplasmic inheritance.
In an area of approximately 400 m², with NB1 biossecurity level, the facilities include: (a) complete structure for in vitro production of embryos in a broad range of domestic species, including equipment for embryo and gamete micromanipulation (composed of inverted epifluorescence microscope equipped with micromanipulators, Narishige microinjectors, FemtoJet® microinjector, electrofusion system and Piezo Drill); as well as embryo and cell in vitro culture systems with controlled atmosphere;

 

 

 

 

 

 

 (b) facilities for cell analysis, composed of flow cytometer (BD FACSaria) with blue (488nm) and near UV (375nm) lasers and cell sorter for immunophenotyping, apoptosis, cell cycle, post-translation protein modification assays; fluorometer for ATP content, calcium and enzymatic activity assays; and epifluorescence microscope for photodocumentation;

 

 

(c) molecular biology facility composed of Real Time PCR (Applied Biosystems 7500 Fast) equipped with High Resolution Melting, Nanodrop 2000, Bioanalyser 2100, thermocyclers, electrophoresis system and 2D gel scanner.

 

 

 

 

The LMMD facilities also include a research laboratory annex to the Veterinary Hospital of FZEA, currently under construction, with an area of 149.5 m² where will be installed the transgenesis, recombinant proteins and cell therapy laboratories. This laboratory will be part of the Experimental Veterinary Hospital dedicated to Innovative Therapies (Unidade Experimental Clínico Hospitalar de Medicina Veterinária) funded by FINEP (Finep Project 01.10.0023.00), that will allow the development of preclinical tests using domestic animals as models.

 

Laboratory for Molecular Genetics

The Laboratory for Molecular Genetics, at the Bioscience Institute, University of Sao Paulo, Works with embryonic stem cells for both basic and applied research. We have established lines of mouse ESCs that are currently used to make mouse models of human genetic diseases. In addition, we work on the establishment of lines of human ESCs in different culture conditions in order to study the epigenetic state of the X chromosome in early human embryonic development.
Finally, our laboratory includes The National Laboratory for Embryonic Stem Cells (LaNCE), a GMP facility for the establishment of new lines of hESCs for therapy.

 

Laboratory of Experimental Models

The group has experience with experimental animal models and immunology. The research lines of the group include: a, Immune mechanisms of action of autologous hematopoietic stem cell transplantation for autoimmune diseases; Treatment of experimental models of autoimmune diseases with mesenchymal stem cell infusions: evaluation of therapeutic response and mechanisms of action; Treatment of extensive burn wounds with mesenchymal stem cells: evaluation of therapeutic response and mechanisms of action; Analysis of functional characteristics and genetic profiling of stem cells isolated from autoimmune patients.