New line of mesenchymal stem cell isolated from warton’s jelly of the umbical cord of male human donor

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Abstract

A new non-immortalized fibroblast-like cell line, named MSCWJ-3, was generated and characterized. Characteristics during long-term cultivation (6–24 passages) confirm the status of MSCs. It is shown: 1) a gradual increase in the proportion of senescent cells during long-term cultivation; 2) a significant decrease in the proliferation index by the 24th passage; 3) preservation of the normal diploid karyotype of the man (46, XY) during the entire period of cultivation, trisomy for different autosomes in single cells, absence of structural chromosomal rearrangements; 4) a high proportion of cells carrying surface antigens characteristic of MSCs: CD44, CD73, CD90, CD105, HLA-ABC and a low proportion with antigens CD34, CD45 and HLA-DR over 24 passages. Cells of the MSCWJ-3 line are capable of differentiation in the osteogenic and adipogenic directions at early and late passages; differentiation in the chondrogenic direction is absent. In general, there are some differences with previously obtained lines isolated from the same source and are associated mainly with the degree of expression of a number of status characteristics.

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About the authors

A. M. Koltsova

Institute of Cytology, Russian Academy of Sciences

Author for correspondence.
Email: koltsova.am@mail.ru
Russian Federation, St. Petersburg

A. S. Musorina

Institute of Cytology, Russian Academy of Sciences

Email: koltsova.am@mail.ru
Russian Federation, St. Petersburg

V. I. Turilova

Institute of Cytology, Russian Academy of Sciences

Email: koltsova.am@mail.ru
Russian Federation, St. Petersburg

A. N. Shatrova

Institute of Cytology, Russian Academy of Sciences

Email: koltsova.am@mail.ru
Russian Federation, St. Petersburg

T. K. Yakovleva

Institute of Cytology, Russian Academy of Sciences

Email: koltsova.am@mail.ru
Russian Federation, St. Petersburg

G. G. Poljanskaya

Institute of Cytology, Russian Academy of Sciences

Email: gpolanskaya@gmail.com
Russian Federation, St. Petersburg

References

Supplementary files

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2. Fig. 1. Morphology of MSCWJ-3 cells at passages 6 and 24. Lifetime photographs. Inverted light microscope Nikon Eclipse TS100 (Japan). Scale bar: 200 µm.

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3. Fig. 2. Growth curves of MSCWJ-3 cell line at passages 6 (curve 1), 16 (curve 2) and 24 (curve 3).

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4. Fig. 3. Karyotype of the MSCWJ-3 cell line. Normal karyotype: 46, XY. The insets show trisomies for different autosomes detected in single cells with 47 chromosomes at passages 6, 16, and 24.

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5. Fig. 4. Identification of markers of early differentiation of human embryonic stem cells in MSCWJ-3 cells at passage 6 by fluorescence of corresponding monoclonal antibodies. Markers of ectoderm (nestin), mesoderm (α-actinin), endoderm (α-fetoprotein) are shown in green. Nuclei are stained with Hoechst 33342. Scale bar: 100 µm.

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6. Fig. 5. Visualization of differentiation of MSCWJ-3 cells in the adipogenic and osteogenic directions at passages 6 and 24. Fat inclusions (adipogenic differentiation) are stained with oil red (scale bar: 100 μm), calcium salts in the intercellular space are revealed by staining with alizarin red and the Von Kossa reaction (scale bar: 200 μm).

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