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Chromosomal reorganization in human embryonic stem cell lines provides a model for cytogenetic research

 

Prokhorovich M.A1., Lagarkova M.A2,4., Karamisheva T.V1., Shilov A.G1., Zhelezova A.I1., Kizilova E.A1., Likhoshvai T.A. 3, Kiselev S.L. 4, Rubtsov N.B1,3.

 

(1 - Institute of Cytology and Genetics SB RAS, Novosibirsk, e-mail: maryalag@yahoo.com)

 

It is well known that chromosome reorganization can influence on the gene expression. In stem cells variation of many different characteristics, including proliferation rate, differentiation potential, response to cytokines, and expression of many gene markers can be associated with karyotype reorganization. Detailed chromosome analysis has to be performed in any study involved human embryonic stem cells (hESCs) due to the possible effect of karyotype reorganization and any chromosomal abnormality revealed in studied cells has to be taken into account in further investigation.

 

Cytogenetic analysis of chromosomes in seven human embryonic stem cell lines (ESM01-04 and their derivatives) was performed with GTG-banding technique. In three sublines of two hESC lines aberrant chromosomes were revealed. Significant differences in proliferation and differentiated properties were observed in hESC lines with chromosomal aberrations. The abnormal karyotypes were stable in these lines during dozens of passages. ESM01r18 cells contained chromosome derived from chromosome 18. Karyotype of this cell line was 46,XX,der(18).

 

For precise description of chromosome der(18) in ESM01r18 line the DNA probe specific to the aberrant chromosome was generated by metaphase chromosome microdissection followed by DOP-PCR. Chromosomal in situ suppression hybridization on metaphase chromosomes from ESM01r18 cells stained, all regions of the der(18). Normal chromosome 18 derived from ESM01r18 cells and from lymphocytes of healthy donors was stained only in p11.31>q21.2 region. No signal was detected on other chromosomes. Analysis of FISH results revealed that the der(18) was a ring chromosome composed of two copies of 18p11.31>q21.2 region located in inverted orientation, r(18)(:: p11.31>q21.2::q21.2>p11.31::). One of two centromears located close to each other was probably inactivated.

 

Additionally to detailed description of the r(18) the comparison of location of normal chromosome 18 and r(18) in interphase nuclei of ESM01r18 cells was performed. For visualization and identification of chromosome territories of the normal chromosome 18 and r(18) two-color FISH with microdissected whole chromosome 18 and DNA probe based on BAC43 was carried out. Due to the loss of the region of BAC43 location during formation of the r(18), the territory of the aberrant chromosome 18 in interphase nuclei differed from chromosome 18 territory by absence of the BAC43 signal. 3D FISH comparative analysis of these two chromosomes location in interphase nuclei revealed the change of spatial position of the aberrant chromosome.

 

Biological significance of genetic alterations in hESC lines has been studied by comparison of in vivo ESM01 and ESM01r18 differentiation abilities. The histological examination of teratomas formed in nude/nude mice revealed differences in teratomas derived from ESM01 and ESM01r18 cells. Overall, chromosome rearrangement analyzed in this study could be associated with variation in proliferation rate and differentiation potential of ESM01r18.

 

 

См. также:

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    The induction of differentation of mouse embryonic stem cells in conditions of prolonged cultivation with lif recombinant protein

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    Possibility of mesenhcymal stem cells application in the treatment of therapy-resistant nervous system disorders

    An influence of mesenchymal stem cells transplantation upon cognitive function restoration after stroke in rats

     ...

 

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Изменен: 13.08.08

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