COMPARATIVE CHARACTERISTICS OF NEW MESENCHYMAL STEM CELL LINES DERIVED
FROM HUMAN EMBRYONIC STEM CELLS, BONE MARROW AND FORESKIN
T.A. Krylova, A.M. Koltsova, V.V. Zenin, A.S. Musorina,
T.K. Yakovleva, G.G. Poljanskaya 1
Institute of Cytology RAS, St. Petersburg;
1 e-mail: poljansk@mail.cytspb.rssi.ru
New nonimmortalized fibroblast-like cell lines SC5-MSC and SC3a-MSC, FetMSC, FRSN were obtained
from human embryonic stem cells (ESC), bone marrow of a 5—6-days embryo and foreskin of a 3-years-old
boy, respectively. All the lines are successfully used as the feeder at human ESC cultivation. It is determined
that the average cel l population doublings time varies from 25.5 h for ISC5-MSC to 38.8 h for SC3a-MSC. Active
proliferation of all the lines is also shown by the corresponding growth curves. Numerical and structural karyotypic
analysis showed that these lines had normal karyotype: 46,XX (SC5-MSC and SC3a-MSC) and 46,XY
(FetMSC and FRSN). To determine the status of the lines, their cell surface markers were analyzed by flow cytometry.
This analysis revealed the presence of surface antigens CD44, CD73, CD90, CD105 and HLA-ABC,
characteristic of human MSC, and the absence of CD34 and HLA-DR. Different lines were found to express
CD117(c-kit) to a different level. Immunofluorescence and flow cytometry analysis did not detect TRA-1-60
and Oct-4, characteristic of human embryonic stem cells, and revealed interlinear variations in the level of
SSEA, which did not depend on the cell origin. It is not clear yet whether these interlinear variations affect functional
MSC status. In all the lines, immunofluorescence analysis showed the presence of the markers of early
differentiation in the derivates of three germ layers which may allow MSC to be useful, in corresponding microenvironments,
for reparation of tissue injures. Adipogenic and osteogenic differentiatiation of all cell lines has
been shown.
Key words: human mesenchymal stem cell lines, immunofluorescence analysis, surface cell markers, karyotype
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