Medical College of Wisconsin
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Generation of mesenchymal stromal cells from HOXB4-expressing human embryonic stem cells. Cytotherapy 2009;11(6):716-25

Date

11/03/2009

Pubmed ID

19878058

Pubmed Central ID

PMC2774218

DOI

10.3109/14653240903051566

Scopus ID

2-s2.0-73649138531 (requires institutional sign-in at Scopus site)   7 Citations

Abstract

BACKGROUND AIMS: HOXB4 transcription factor plays an important role in embryonic and adult hematopoiesis. Overexpression of HOXB4 in murine and human embryonic stem cells (ESC) has been used to generate hematopoietic stem cells (HSC) via the embryoid body formation method.

METHODS: We used FuGENE 6-based transfection of YPL2-HOXB4 vector to generate HOXB4-expressing colonies from human ESC line H9 and investigated the potential of these cells for differentiation into primitive CD34(+) hematopoietic cells, via co-culture methodology with OP9 murine bone marrow stromal cells. Expression of HOXB4 in transfected human ESC colonies and their derivatives was verified using immunocytochemistry and reverse-transcription polymerase chain reaction (RT-PCR).

RESULTS: Utilizing OP9 stromal cell co-culture methodology, we generated CD34(+) cells from HOXB4-expressing H9 human ESC at a frequency similar to, and not higher than, non-transfected human ESC. However, we observed that some colonies of HOXB4-expressing human ESC not co-cultured on OP9 cells, differentiated into mesenchymal stromal cells (MSC) while preserving their HOXB4 expression. These HOXB4-expressing MSC expressed CD29, CD73, CD44, CD90, CD105 and HLA-class I, were negative for the expression of CD34, CD45, CD54, CD71, CD106 and HLA-DR, and could be differentiated into adipocytes and osteocytes.

CONCLUSIONS: In our specific experimental system we observed that overexpression of HOXB4 in human ESC did not improve the generation of CD34(+) hematopoietic cells via OP9 co-culture methodology. Furthermore, we could generate MSC from human ESC over-expressing HOXB4.

Author List

Liu YP, Hematti P

Author

Peiman Hematti MD Professor in the Medicine department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Antigens, CD
Cell Differentiation
Cell Line
Coculture Techniques
Embryonic Stem Cells
Homeodomain Proteins
Humans
Mice
Stromal Cells
Transcription Factors
Transfection