In vitro expression of erythropoietin by transfected human mesenchymal stromal cells.
BACKGROUND
Mesenchymal stromal cells (MSC) are pluripotent progenitor cells that can be found in human bone marrow (BM). These cells have low immunogenicity and could suppress alloreactive T-cell responses. In the current study, MSC were tested for their capacity to carry and deliver the erythropoietin (EPO) gene in vitro.


METHODS
Expanded BM MSC was transfected with EPO-encoded plasmid pMCV1.2 and EPO-encoded MIDGE (minimalistic immunologically defined gene expression) vector by electroporation. The expressed EPO was used to induce hematopoietic stem cells (HSC) into erythroid colonies.


RESULTS
The results showed that the MIDGE vector was more effective and stable than the plasmid (pMCV1.2) in delivering EPO gene into MSC. The supernatants containing EPO obtained from the transfected cell culture were able to induce the differentiation of HSC into erythroid colonies.


CONCLUSIONS
MSC hold promise as a cell factory for the production of biologic molecules, and MIDGE vector is more effective and stable than the plasmid in nucleofection involving the EPO gene.
Mok
P-L
PL
Cellular Therapy Unit, MAKNA-HUKM Cancer Institute, Kuala Lumpur, Malaysia.
Cheong
S-K
SK
Leong
C-F
CF
Othman
A
A
D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D003114:Colony-Forming Units Assay; D041905:Erythroid Cells / Q000166:cytology; D004921:Erythropoietin / Q000235:genetics*; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D008648:Mesoderm / Q000166:cytology*; D010957:Plasmids / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D014162:Transfection
D004921:Erythropoietin