Obtaintion of erythrocytes from induced pluripotent stem cells in feeder-free culture with xeno-free culture medium

• Grant number: 16/08373-9
• Support type: Scholarships in Brazil - Master
• Effective date (Start): July 01, 2016
• Effective date (End): June 30, 2018
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Virginia Picanço e Castro
• Grantee: Bárbara Cristina Martins Fernandes Paes
• Home 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 research grant: 13/08135-2 - CTC - Center for Cell-Based Therapy, AP.CEPID

Abstract

Hematopoiesis is the process of formation of blood cellular components from the differentiation of hematopoietic stem cells (HSC). The HSCs are found in adult human bone marrow cells at a ratio of 1 in every 10^6 cells. The difficulty in obtaining these cells and determining a protocol for ex vivo expansion leads us to seek alternative sources for the production of mature blood cells. Induced pluripotent stem cells (iPS) are able to differentiate into all cell types of the adult body. Thus, induction of differentiation of iPS cells into hematopoietic cells is an alternative way for the production of erythrocytes in vitro. The production of red blood cells in vitro is needed for the study of diseases, development of therapies and clinical applications involving this cell type. However, for such cells to have potential clinical applications it is necessary that, among other requirements, they are cultured without feeder cells and animal molecules. This project proposes the production of red blood cells from induced pluripotent stem cells in culture free of feeder cells and in medium free of animal molecules. The hematopoietic differentiation will follow the embryoid body formation method, and subsequent induction of erythroid differentiation. During differentiation of iPS cells, erythroid and hematopoietic development will be monitored by colony formation analysis and flow cytometry analysis. The expression of genes related to hemoglobins will be checked by real time RT-PCR. Analysis of hemoglobin composition in the produced erythrocytes will be performed by cation exchange high efficienty liquid chromatography (HPLC).

Scientific publications

(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)

MARTINS FERNANDES PAES, BARBARA CRISTINA; MOCO, PABLO DIEGO; PEREIRA, CRISTIANO GONCALVES; PORTO, GECIANE SILVEIRA; DE SOUSA RUSSO, ELISA MARIA; JUNQUEIRA REIS, LUIZA CUNHA; COVAS, DIMAS TADEU; PICANCO-CASTRO, VIRGINIA. Ten years of iPSC: clinical potential and advances in vitro hematopoietic differentiation. CELL BIOLOGY AND TOXICOLOGY, v. 33, n. 3, p. 233-250, MAY 2017. Web of Science Citations: 6.