502-509Mesenchymal stromal cells (MSC) are
multipotent cells that can be derived from many different organs and tissues.
While there are many ways to label and track cells each with strengths and
weakness, the green fluorescent protein (GFP) is a reporter gene commonly
employed. In the present study, caprine MSC were collected from bone marrow and
cells were characterised with MSC specific markers. Passage 10 (P10) MSC cells
were transfected using plasmid vector containing GFP as reporter gene with
different concentrations of DNA and lipofectamine. Six different concentrations
of DNA and lipofectamine as 1 µg DNA: 2 µL lipofectamine, 1 µg DNA: 2.5 µL
lipofectamine, 1.2 µg DNA: 2.2 µL lipofectamine,
1.2 µg DNA: 2.5 µL lipofectamine, 1.5 µg DNA: 2.5 µL lipofectamine, 1.5 µg DNA:
3 µL lipofectamine were used. After 24 h and 48 h of transfection, caprine MSC
were observed under florescent microscope. Highest transfection rate indicating
green flourecscent MSC were found when the cells were transfected with 1.2 µg
DNA: 2.2 µL lipofectamine and 1.5 µg DNA: 2.5 µL lipofectamine than other
combinations. These cells have been propagated beyond 4th passage
maintaining GFP expression. The results indicated that stable GFP positive MSC
cells can be generated using the above protocol. These cells are being used for
transplantation studies