Experimental Protocols with Animal Models and IVIS Spectrum (In Vivo Imaging System)

Core Facilities

Center for Cell-Based Therapy maintains a variety of several core facilities to support ours research programs and educational activities, including laboratories of Cell Therapy, Molecular Biology, Gene Transfer, Molecular Genetic Analysis, Flow Cytometry, Bioinformatics, Protein Chemistry, and HLA Genotyping. The CTC core facilities are staffed by highly experienced researchers in the technologies offered.

Cell Therapy Core Confocal and Microscopy DNA sequencing
Domestic Animal Facility Embryo Micromanipulation Flow Cytometry
Experimental Hematology HLA Genotyping Library
Mass Spectrometry GMP facility for cell culture  
High Content Screening (HCS)

Experimental Protocols with Animal Models 

IVIS Lumina (In Vivo Imaging System)

 

 

 

Experimental Hematology Laboratory

The Experimental Hematology Laboratory works at the Hospital of Ribeirão Preto Medical School (USP) and the Blood Center of Ribeirão Preto, working in the diagnostics and research in blood diseases.
     
 

Flow Cytometry Lab

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. In order to follow the technological advances, the Hemocentro has recently bought the Flow Cytometer, which is able to perform the “cell sorter” (separation of subpopulation), to perform simultaneous analyses on seven different fluorescence, and so forth.

Principal Investigator: Prof. Dr. Dimas Tadeu Covas

Manager: Patricia Vianna Bonini Palma

 

     
  Cell Therapy Core Facility

Our goal is to provide Hematopoietic (HSCs) and Mesenchymal stem cells (MSCs) for clinical researchers performed by the Center for Cell-Based Therapy.  We have the expertise to manipulate and cryopreserve HSCs for transplantation since 1997. In our facility, the products which contain hematopoietic stem cell (HSC) such as bone marrow, umbilical cord blood and mobilized peripheral blood are processed in order to keep their biological function until the moment of usage, which can happen years after the collection. We also expanded MSC in relative large scale in culture system using T flask. The MSCs are tested for safety (sterility, mycoplasma, adventitious virus), identity/purity (morphology, antigenic phenotype and cell viability by flow cytometry), potency to inhibit T lymphocyte proliferation and to differentiate into adipocytes and osteocytes, purity (endotoxin) and stability (consistent composition, stable karyotype [G-Banding] and ability to differentiate and to inhibit T lymphocyte proliferation) after long term culture. Our next step is to create allogeneic MSC cell bank, where the cryopreserved banked cells will be previously tested, thus ready for “off-the-shelf” clinical use.

Principal Investigator: Prof. Dr. Dimas Tadeu Covas

Managers: Dra. Maristela Delgado Orellana and Dr. Gil Cunha de Santis

    

Cell Culture Core Facility

Our goal is to provide technical support to Center for Cell-Based Therapy researchers in projects to adult and embryonic stem cells. We also conduct basic research with human pluripotent and multipotent stem cells. Our major focus is the generation, in vitro, of hematopoietic cells and endothelial from human embryonic (hESCs) and induced pluripotent stem cells (hiPS). These studies are used to characterize molecular and cellular mechanism underpinning the differentiation of hematopoietic progenitors and mature hematopoietic cells. This research will contribute to clinical translation of hESC- and iPSC-derived hematopoietic cells for novel therapies in the next few years. The laboratory also performs researches on multipotent mesenchymal stromal cells (MSCs). These studies comprise MSCs isolation, culture, expansion and characterization, contributing for a better understanding of the potential clinical usage of these cells.

Principal Investigator: Prof. Dr. Dimas Tadeu Covas

Manager: Dra. Maristela Delgado Orellana

      

     
  Mass Spectrometry and Protein Chemistry Lab

The Protein Chemistry Center (CQP) provides scientific and technical support for proteomics and mass spectrometry analysis. The Center for proteomics and mass spectrometry is equipped with three mass spectrometer (ESI-Triple Quadrupole, ESI-Q-TOF and MALDI-TOF/TOF) and complete systems for on line and off line protein/peptide separation by capillary liquid chromatography, specialized software for protein quantification and identification using local or internet databases.

Principal Investigator: Prof. Dr. Lewis Joel Greene

 

     
  Bioinformatics Team

The Bioinformatic Team (BiT) provides scientific and technical support for molecular biology, genomics, proteomics and other biological related approaches. The BiT Lab is equipped with high-performance computer server, specialized software, and advanced database systems to genomic analysis.

Principal Investigator: Prof. Dr. Wilson A. Silva Jr.

Manager: Raul Torrieri  

     
  DNA sequencing and Next Generation Sequencing Lab

The High-Throughput DNA and RNA Sequencing facility provides access to automated DNA sequencing technology. The services offered include experimental design consultation, library preparation, data export, and primary and secondary data analysis.

Next Generation Sequencing (NGS)

For the investigation of genetic and epigenetic changes. The epigenomic signatures are obtained by large-scale or new-generation sequencing. The experience of the group with the analysis of data generated by large-scale sequencing has permitted the development of a new bioinformatic tool called biOMICS (Biologically Integrating Omics), whose objective is to integrate information from the projects ENCODE, NIH Roadmap and The Cancer Genome Atlas (TCGA) with new large-scale sequencing data. These activities are coordinated by Houtan Noushmehr, a collaborator linked to the group coordinated by Wilson Araújo da Silva Junior (PI).

Principal Investigator: Prof. Dr. Wilson A. Silva Jr.

Managers: Adriana Aparecida Marques and Kamila Peroni

     
 

Confocal and Microscopy

Confocal Microscopy Laboratory is a facility designed to serve as a service center for researchers in need of primary stem cells, tumor cells from either human or murine analyses. We are specialized in morphology characterization, detection of specific proteins by immunofluorescence and detection of fluorescence-labelled cells. We study cultured normal and tumor cells and tissue samples from either human or animal models. The core is equipped with Laser Scanning Microscopy (LSM710, Carl Zeiss).

Principal Investigator: Prof. Dr. Dimas Tadeu Covas

Manager: Patricia Vianna Bonini Palma

 

     
 

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 the São Paulo State; 2, living kidney and bone marrow donors (family members) for transplantation in the University Hospital of School of Medicine 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”.

Principal Investigator: Prof. Dr. Eduardo Antônio Donadi

Manager: Neifi Hassan Saloum Deghaide

     
  Embryo Micromanipulation

The Embryo Micromanipulation lab have a complete structure for in vitro production of embyos in a broad range of domestic species, including equipments for embryo and gamete micromanipulation (compose of inverted epifluorescence microscope equipped with micromanipulators, Narishige microinjectors, FemtoJet® microinjector, electrofusion system, Piezo Drill, besides the equipment’s to produce borosilicate tools); as well as embryo and cell in vitro culture systems with controlled atmosphere.

Principal Investigator: Prof. Dr. Flávio V. Meirelles

Manager:  Dra. Fabiana Fernandes Bressan

     
  Domestic Animal Facility

This facility is an experimental Animal Hospital expected to be ready for use on July 2012, created to serve as a Laboratory for teaching and for doing research on innovative therapies (INOVATERAP) financed by FINEP.

This experimental facility will have 3,000 m2 infrastructure for large and small domestic animal care, clinic, image and lab. analysis, as well as surgery and pathology. Besides of the hospital facility a 143 m2 lab. will be available to produce and handle cells Germ free condition.

Principal Investigator: Prof. Dr. Flávio V. Meirelles

Manager: Dra. Fabiana Fernandes Bressan

     
 

GMP facility for cell culture

The GMP cell culture facility can be used for the production of cell-based products for therapy.  It contains an XVivo isolator, where cell handling is done without any contact with the exterior.

Principal Investigator: Profa. Dra. Lygia V. Pereira

Manager: Dra. Mariana Marques Morato

     
 

High Content Screening (HCS) 

High-Content Screening (HCS) refers to a group of experimental approaches that consists in the use of multi-well imaging plates (96 to 384-well plates) to carry cell-based functional assays, allowing multiple treatment conditions to be evaluated simultaneously. The HCS assays are based on the use of fluorescent reagents and automated immunofluorescence microscopy. The automated acquisition of digital images, coupled to their automated processing and analysis, allows diverse molecular and morphological events to be quantified at a single-cell-level.

The high-content of these multiparametric readouts, allow diverse complex biological processes to be interrogated simultaneously, following perturbations with different chemical or biological compounds (e.g. unknown or known drug inhibitors or agonists, cytokines, growth factors, etc) or genetic tools (siRNA, esiRNAs, shRNAs, microRNA mimics, microRNA inhibitors, CRISPR sgRNAs, etc).

The LLSFBio provides support for all phases involved in a HCS project, including: 1) project evaluation and technical consultation, 2) HCS assay development and validation, 3) lab automation and screening execution, 4) automated image acquisition, 5) quantitative image analysis and 6) Data analysis.

More information.

Principal Investigator: Prof. Dr. Dimas Tadeu Covas 

Manager: Dr. Rodrigo Alexandre Paneppuci

  

     
 

Experimental Protocols with Animal Models 

 

The differential of the laboratory is a room availability for the execution of the experimental protocols required by the researchers of the institution and external collaborators, facilitating logistics related to the experiments. We have equipment such as laminar flows, exchange booths, inhalation anesthesia machines, centrifuges, freezers, refrigerators, microscopes, incubators, water baths and fluorescent and bioluminescent imaging equipment in vivo (IVIS® Lumina).

Principal Investigator: Prof. Dr. Eduargo Magalhães Rego

Manager: Ma. Cleide Lúcia Araújo da Silva

 

     
 

IVIS Lumina (In Vivo Imaging System) 

 

 

The Xenogen IVIS® Lumina equipment has a high sensitivity technology that allows non-invasive visualization and tracking of cellular and genetic activity using live organisms in real time when there is integration into the cells/tissues of fluorescent or bioluminescent reporter components (luciferase).

Principal Investigator: Prof. Dr. Eduargo Magalhães Rego

Manager: Ma. Cleide Lúcia Araújo da Silva

  

     
 

Library (42 m²)

This unit have 2,574 books, 91 titles of periodicals and journals,  375 thesis and dissertations, guides, CAPES electronic signatures, 8 journals signatures, annals and collection of scientific production of local researchers.

Manager: Marjory Cristina Donato

 

 

 

 

 

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