Derivation of normal macrophages from human embryonic stem (hES) cells for applications in HIV gene therapy.

Retrovirology
Joseph S AndersonRamesh Akkina

Abstract

Many novel studies and therapies are possible with the use of human embryonic stem cells (hES cells) and their differentiated cell progeny. The hES cell derived CD34 hematopoietic stem cells can be potentially used for many gene therapy applications. Here we evaluated the capacity of hES cell derived CD34 cells to give rise to normal macrophages as a first step towards using these cells in viral infection studies and in developing novel stem cell based gene therapy strategies for AIDS. Undifferentiated normal and lentiviral vector transduced hES cells were cultured on S17 mouse bone marrow stromal cell layers to derive CD34 hematopoietic progenitor cells. The differentiated CD34 cells isolated from cystic bodies were further cultured in cytokine media to derive macrophages. Phenotypic and functional analyses were carried out to compare these with that of fetal liver CD34 cell derived macrophages. As assessed by FACS analysis, the hES-CD34 cell derived macrophages displayed characteristic cell surface markers CD14, CD4, CCR5, CXCR4, and HLA-DR suggesting a normal phenotype. Tests evaluating phagocytosis, upregulation of the costimulatory molecule B7.1, and cytokine secretion in response to LPS stimulation showed that these macro...Continue Reading

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Aug 22, 2013·Stem Cells and Development·Anne TannerBradford K Berges
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Aug 31, 2021·Stem Cells·Somayeh PouyanfardDan S Kaufman

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Methods Mentioned

BETA
transgenic
FACS
ELISA
scraping

Software Mentioned

EXPO32 ADC

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