Visit our Expo - Redox and Inflammation signaling 2012

VII. Stem cell specific cell signaling mechanisms Poster VII, 1
Murine marrow-derived mesenchymal stem cells:Isolation,Characterisation
*Raza Haji Hosseini ,**Samad Nadri
*Biology Dept. Faculty of science, Payame Noor University, Tehran, Iran 19569
*E.mail:hosseini@pnu.ac.ir,
**Biology Dept. Faculty of science, Payame Noor University, Tehran, Iran 19569
**E.mail:nadrisamad@yahoo.co.uk
Introduction: Many researchers have employed different methods to isolate and purify
mesenchymal stem cells in mouse bone marrow but there is a lack of consensus on
mesenchymal stem cells express surface markers. There is evidence for the affect cell density
on surface markers in human mesenchymal stem cells. IN present study, Expression of
CD34,CD44,Sca-1,c-Kit,Vcam-1 on MSCs purified by Low density and High density from
NMRI and BALB/c was studied.
Method and Material:6-8 week-old NMRI andBALB/c were killed by cervical dislocation and
bone marrow were flushed and cultured using low glucose DMEM medium by Low density
and High density methods then MSCs were purified by continue passages.To examine the
mesenchymal nature of isolated cells,The cells were differentiated into bone and
adipocyte.the cells were abeled by CD34,CD44,Sca-1,c-Kit,Vcam-1 antibody conjugated to
PE or FITC and analyzed by flow cytometry method using FACS-analysis Machine.
Result:Spindle –shaped purified cells were observed in Low density and High density cultures
at the end of the fourth and seven passage,respectively.The cells from both NMRI
andBALB/c were easily differentiated into osteogenic and adipogenic Lineages.Flow
cytometry analysis indicated that The MSCs from either NMRI and BALB/c mice are CD44+
,Sca-1+ CD34- and c-Kit- in Low density and High density culture.The mMSCs from tow
strains differed in their expression of the cell-surface epitope Vcam-1.
Conclusion:In present investigation,The presence of five surface markers was investigated on
MSCs purified by Low density and High density culture from NMRI and BALB/c.The results
presented here,indicated up to 90% cells is CD44+ , It seems That It could be useful markers
in isolation of the MSCs.Comparisons of MSCs isolated from 2 strains demonstrated That
some features of The cells were strain specific also They isolation more rapidly if plated at
Low density culture. In this studies, In spite of previously research,Cell density is not affected
on expression surface markers.
Key Word:marker,mesenchymal stem cell,bone marrow
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