Hematopoietic Stem Cells transfection with retrovirus using ViroMag

The LSK cell fraction of HSC was transduced with retrovirus on retronectin using ViroMag.

The paper shows the high efficiency of Magnetofection method from OZ Biosciences to improve infection and transduction capacity on hematopoietic stem cells.

article reference: Biochem Biophys Res Commun. 2013 Oct 16.

Nucleostemin is indispensable for the maintenance and genetic stability of hematopoietic stem cells.

Yamashita M, Nitta E, Nagamatsu G, Ikushima YM, Hosokawa K, Arai F, Suda T.

Abstract
Nucleostemin is a nucleolar protein known to play a variety of roles in cell-cycle progression, apoptosis inhibition, and DNA damage protection in embryonic stem cells and tissue stem cells. However, the role of nucleostemin in hematopoietic stem cells (HSCs) is yet to be determined. Here, we identified an indispensable role of nucleostemin in mouse HSCs. Depletion of nucleostemin using short hairpin RNA strikingly impaired the self-renewal activity of HSCs both in vitro and in vivo. Consistently, loss of nucleostemin triggered apoptosis rather than cell-cycle arrest in HSCs. Furthermore, DNA damage accumulated during cultivation upon depletion of nucleostemin. The impaired self-renewal activity of HSCs induced by nucleostemin depletion was partially rescued by p53 deficiency but not by p16^Ink4a or p19^Arf deficiency. Taken together, our study demonstrates that nucleostemin protects HSCs from DNA damage accumulation and is required for the maintenance of HSCs.

ViroMag from OZ Biosciences is a magnetic nanoparticles formulation optimized for increasing any kind of virus infection and transduction both in vitro and in vivo.