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In vitro expansion of mesenchymal stromal cells and secretome characterization: therapeutic and biotechnological applications

Grant number: 12/23228-4
Support Opportunities:Scholarships in Brazil - Doctorate
Effective date (Start): March 01, 2013
Effective date (End): May 31, 2016
Field of knowledge:Health Sciences - Medicine - Medical Clinics
Principal Investigator:Kamilla Swiech Antonietto
Grantee:Amanda Mizukami Martins
Host Institution: Hemocentro de Ribeirão Preto. Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto da USP (HCMRP). Secretaria da Saúde (São Paulo - Estado). Ribeirão Preto , SP, Brazil
Associated scholarship(s):13/23599-5 - Expansion of mesenchymal stromal cells derived from human umbilical cord matrix in bioreactors in serum/xeno-free conditions, BE.EP.DR

Abstract

The mesenchymal stromal cells (MSCs) have become of great interest for cell therapy because of its potential to differentiate and replenish specialized tissues. More recently, such interest has increased significantly due to the discovery that MSCs are capable of secreting a multitude of mediators to stimulate the regeneration of injured tissues in situ and immunomodulators. Therefore, MSCs may be considered both as a therapeutic product itself, such as a biofactory various relevant proteins from the standpoint of therapeutic and biotechnological. To meet these growing demands, both applications require the development of high-throughput processes under defined culture conditions, reproducible, cost-effective, allowing to obtain products with appropriate identity, strength, purity and safety. Therefore, the principal objectives of this study are to establish an efficient expansion process both in terms of growth and recovery of mesenchymal stromal cells and characterize the secretome, or the complement of proteins secreted by these cells during expansion, in order identifying possible relevant proteins with therapeutic applications. For the development of an efficient expansion process will evaluate many types of bioreactors (stirred tank, fixed bed, hollow-fibers). The growth and cell metabolism will be identified during the expansion process. The analysis of the maintenance of the biological properties (inhibition of proliferation and differentiation of lymphocytes) and immunophenotypic cells will be evaluated after cell recovery. To characterize quantitatively and the set of proteins secreted by MSCs during the expansion process, a quantitative approach will be used in marking cell culture with stable isotopes (SILAC), together with fractionation of intact proteins and analysis by mass spectrometry High resolution liquid chromatography coupled to (LC-MS/MS). Thus, by analyzing the various types of bioreactors is expected to introduce an efficient expansion of MSCs with high recovery functional and enabling its wide therapeutic applicability and identify secreted proteins involved in important trophic function of MSCs in cellular therapy and which may represent an interesting by-product of these cells in vitro.

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Scientific publications (10)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
TOZETTI, PATRICIA APARECIDA; CARUSO, SAMIA RIGOTTO; MIZUKAMI, AMANDA; FERNANDES, TAISA RISQUE; DA SILVA, FERNANDA BORGES; TRAINA, FABIOLA; COVAS, DIMAS TADEU; ORELLANA, MARISTELA DELGADO; SWIECH, KAMILLA. Expansion Strategies for Human Mesenchymal Stromal Cells Culture under Xeno-Free Conditions. BIOTECHNOLOGY PROGRESS, v. 33, n. 5, p. 1358-1367, . (13/08135-2, 12/23228-4)
MIZUKAMI, AMANDA; FERNANDES-PLATZGUMMER, ANA; CARMELO, JOANA G.; SWIECH, KAMILLA; COVAS, DIMAS T.; CABRAL, JOAQUIM M. S.; DA SILVA, CLAUDIA L.. Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells. Biotechnology Journal, v. 11, n. 8, p. 1048-1059, . (13/08135-2, 12/23228-4, 13/23599-5)
MIZUKAMI, AMANDA; THOME, CAROLINA HASSIBE; FERREIRA, GERMANO AGUIAR; LANFREDI, GUILHERME PAUPERIO; COVAS, DIMAS TADEU; PITTERI, SHARON J.; SWIECH, KAMILLA; FACA, VITOR MARCEL. Proteomic Identification and Time-Course Monitoring of Secreted Proteins During Expansion of Human Mesenchymal Stem/Stromal in Stirred-Tank Bioreactor. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 7, . (13/08135-2, 12/23228-4)
MIZUKAMI, AMANDA; DE ABREU NETO, MARIO SOARES; MOREIRA, FRANCISCO; FERNANDES-PLATZGUMMER, ANA; HUANG, YI-FENG; MILLIGAN, WILLIAM; CABRAL, JOAQUIM M. S.; DA SILVA, CLAUDIA L.; COVAS, DIMAS T.; SWIECH, KAMILLA. A Fully-Closed and Automated Hollow Fiber Bioreactor for Clinical-Grade Manufacturing of Human Mesenchymal Stem/Stromal Cells. STEM CELL REVIEWS AND REPORTS, v. 14, n. 1, p. 141-143, . (13/08135-2, 12/23228-4)
GONZALEZ GIL, LISETH VIVIANA; SINGH, HARMINDER; DA SILVA, JULIANA DE SA; DOS SANTOS, DIOGO PERES; COVAS, DIMAS TADEU; SWIECH, KAMILLA; TORRES SUAZO, CLAUDIO ALBERTO. Feasibility of the taylor vortex flow bioreactor for mesenchymal stromal cell expansion on microcarriers. Biochemical Engineering Journal, v. 162, . (12/23228-4, 13/08135-2)
PICANCO-CASTRO, VIRGINIA; MOCO, PABLO DIEGO; MIZUKAMI, AMANDA; VAZ, LETICIA DELFINI; FERNANDES PEREIRA, MARCELO DE SOUZA; SILVESTRE, RENATA NACASAKI; COTTAS DE AZEVEDO, JULIA TEIXEIRA; BOMFIM, ALINE DE SOUSA; DE ABREU NETO, MARIO SOARES; RIBEIRO MALMEGRIM, KELEN CRISTINA; et al. Establishment of a simple and efficient platform for car-t cell generation and expansion: from lentiviral production to in vivo studies. Hematology, Transfusion and Cell Therapy, v. 42, n. 2, p. 150-158, . (17/09491-8, 16/08374-5, 13/08135-2, 12/23228-4)
MIZUKAMI, AMANDA; SWIECH, KAMILLA. Mesenchymal Stromal Cells: From Discovery to Manufacturing and Commercialization. STEM CELLS INTERNATIONAL, . (13/08135-2, 12/23228-4)
DA SILVA, JULIANA DE SA; MIZUKAMI, AMANDA; GONZALEZ GIL, V, LISETH; DE CAMPOSA, JESSICA VALERIA; ASSIS, ODILIO B. G.; COVAS, DIMAS TADEU; SWIECHB, KAMILLA; TORRES SUAZO, CLAUDIO ALBERTO. Improving wave-induced motion bioreactor performance for human mesenchymal stromal cell expansion. Process Biochemistry, v. 84, p. 143-152, . (12/23228-4, 13/08135-2)
MOREIRA, FRANCISCO; MIZUKAMI, AMANDA; DE SOUZA, LUCAS EDUARDO BOTELHO; CABRAL, JOAQUIM M. S.; DA SILVA, CLAUDIA L.; COVAS, DIMAS T.; SWIECH, KAMILLA. Successful Use of Human AB Serum to Support the Expansion of Adipose Tissue-Derived Mesenchymal Stem/Stromal Cell in a Microcarrier-Based Platform. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 8, . (13/08135-2, 12/23228-4)
MIZUKAMI, AMANDA; PEREIRA CHILIMA, TANIA D.; ORELLANA, MARISTELA D.; ABREU NETO, MARIO; COVAS, DIMAS T.; FARID, SUZANNE S.; SWIECH, KAMILLA. Technologies for large-scale umbilical cord-derived MSC expansion: Experimental performance and cost of goods analysis. Biochemical Engineering Journal, v. 135, p. 36-48, . (13/23599-5, 13/08135-2, 12/23228-4)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
MARTINS, Amanda Mizukami. Expansion in vitro of mesenchymal stem cell and secretome characterization: therapeutic and biotechnology applications. 2016. Doctoral Thesis - Universidade de São Paulo (USP). Faculdade de Medicina de Ribeirão Preto (PCARP/BC) Ribeirão Preto.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.