DOI: 10.5176/978-981-08-8669-1_SCR2011_5

Authors: Ruzanna Ab Kadir, Shahrul Hisham Zainal Ariffin, Sahidan Senafi, Nazaruddin Ramli, Rohaya Megat Abdul Wahab

Abstract:
Peripheral blood mononucleated cells are consist of leukocytes as well as haematopoietic stem cells (HSC) and mesenchymal stem cells. However, the method obtaining HSC and MSC from peripheral blood mononucleated cells are still debated. In this preliminary study, our aim was to isolate and characterize mononucleated cells from human peripheral blood. Peripheral blood mononucleated cells were isolated using the Ficoll-Paque density-gradient separation method. After 4 days of culture, adherent and suspension mononucleated cells were separated and cultured until 14 days in vitro in media selection. Reverse transcriptase polymerase chain reaction (RTPCR) was used to identify and characterize the expression of stem cell factor receptor (KIT), a HSC marker (signalling lymphocytic activation molecule family member 1, SLAMF1) and a MSC marker (CD105). These revealed that adherent mononucleated cells were positive for MSC markers whereas, mononucleated cells in suspension were positive for HSC
markers. These studies demonstrate the possibility to obtain HSC and MSC population from peripheral blood. Our approach may lead to consideration about improving method of isolating HSC as well as MSC from peripheral blood mononucleated cells and it can be a basis for further research in this filed.

Keywords: mononucleated cells, peripheral blood, suspension cells, adherent cells

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