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Similar biological characteristics of human embryonic stem cell lines with normal and abnormal karyotypes.

Sun X, Long X, Yin Y, Jiang Y, Chen X, Liu W, Zhang W, Du H, Li S, Zheng Y, Kong S, Pang Q, Shi Y, Huang Y, Huang S, Liao B, Xiao G, Wang W - Hum. Reprod. (2008)

Bottom Line: Their biological characteristics, short tandem repeat loci, HLA typing, differentiation capability and imprinted gene, DNA methylation and X chromosome inactivation status were compared between different cell lines.Imprinted gene expression and DNA methylation were also similar among these cell lines.Non-random X chromosome inactivation patterns were found in XX cell lines.

View Article: PubMed Central - PubMed

Affiliation: Institute of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, People's Republic of China. xiaofangsun@hotmail.com

ABSTRACT

Background: Human embryonic stem cell (hESC) lines derived from poor quality embryos usually have either normal or abnormal karyotypes. However, it is still unclear whether their biological characteristics are similar.

Methods: Seven new hESC lines were established using discarded embryos. Five cell lines had normal karyotype, one was with an unbalanced Robertsonian translocation and one had a triploid karyotype. Their biological characteristics, short tandem repeat loci, HLA typing, differentiation capability and imprinted gene, DNA methylation and X chromosome inactivation status were compared between different cell lines.

Results: All seven hESC lines had similar biological characteristics regardless of karyotype (five normal and two abnormal), such as expression of stage-specific embryonic antigen (SSEA)-4, tumor-rejection antigen (TRA)-1-81 and TRA-1-60 proteins, transcription factor octamer binding protein 4 mRNA, no detectable expression of SSEA-1 protein and high levels of alkaline phosphatase activity. All cell lines were able to undergo differentiation. Imprinted gene expression and DNA methylation were also similar among these cell lines. Non-random X chromosome inactivation patterns were found in XX cell lines.

Conclusions: The present results suggest that hESC lines with abnormal karyotype are also useful experimental materials for cell therapy, developmental biology and genetic research.

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Related in: MedlinePlus

Octamer binding protein 4 expression by RT–PCR.Lanes 3–9 represent FY-hES-1, -3, -4, -5, -7, -8 and FY-3PN, respectively. Lane 1 is a negative control and lane 2 is human β-actin (265 bp).
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DEN137F2: Octamer binding protein 4 expression by RT–PCR.Lanes 3–9 represent FY-hES-1, -3, -4, -5, -7, -8 and FY-3PN, respectively. Lane 1 is a negative control and lane 2 is human β-actin (265 bp).

Mentions: Cells in all seven lines showed a high level of AP activity and strongly expressed TRA-1-60, TRA-1-81 and SSEA-4 (Fig. 1II, A, D, E and C, respectively) but not SSEA-1 proteins (Fig. 1II, B). Oct-4 mRNA expression was observed in all seven hESC lines (Fig. 2). Sixteen STR loci were analyzed for hESC lines and each cell line showed distinct STR loci indicating that they were derived from different embryos (Fig. 3A–G). HLA typing also showed that the seven lines have different HLA-A and DBR loci (Table III).


Similar biological characteristics of human embryonic stem cell lines with normal and abnormal karyotypes.

Sun X, Long X, Yin Y, Jiang Y, Chen X, Liu W, Zhang W, Du H, Li S, Zheng Y, Kong S, Pang Q, Shi Y, Huang Y, Huang S, Liao B, Xiao G, Wang W - Hum. Reprod. (2008)

Octamer binding protein 4 expression by RT–PCR.Lanes 3–9 represent FY-hES-1, -3, -4, -5, -7, -8 and FY-3PN, respectively. Lane 1 is a negative control and lane 2 is human β-actin (265 bp).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2538585&req=5

DEN137F2: Octamer binding protein 4 expression by RT–PCR.Lanes 3–9 represent FY-hES-1, -3, -4, -5, -7, -8 and FY-3PN, respectively. Lane 1 is a negative control and lane 2 is human β-actin (265 bp).
Mentions: Cells in all seven lines showed a high level of AP activity and strongly expressed TRA-1-60, TRA-1-81 and SSEA-4 (Fig. 1II, A, D, E and C, respectively) but not SSEA-1 proteins (Fig. 1II, B). Oct-4 mRNA expression was observed in all seven hESC lines (Fig. 2). Sixteen STR loci were analyzed for hESC lines and each cell line showed distinct STR loci indicating that they were derived from different embryos (Fig. 3A–G). HLA typing also showed that the seven lines have different HLA-A and DBR loci (Table III).

Bottom Line: Their biological characteristics, short tandem repeat loci, HLA typing, differentiation capability and imprinted gene, DNA methylation and X chromosome inactivation status were compared between different cell lines.Imprinted gene expression and DNA methylation were also similar among these cell lines.Non-random X chromosome inactivation patterns were found in XX cell lines.

View Article: PubMed Central - PubMed

Affiliation: Institute of Gynecology and Obstetrics, The Third Affiliated Hospital of Guangzhou Medical College, Guangzhou, People's Republic of China. xiaofangsun@hotmail.com

ABSTRACT

Background: Human embryonic stem cell (hESC) lines derived from poor quality embryos usually have either normal or abnormal karyotypes. However, it is still unclear whether their biological characteristics are similar.

Methods: Seven new hESC lines were established using discarded embryos. Five cell lines had normal karyotype, one was with an unbalanced Robertsonian translocation and one had a triploid karyotype. Their biological characteristics, short tandem repeat loci, HLA typing, differentiation capability and imprinted gene, DNA methylation and X chromosome inactivation status were compared between different cell lines.

Results: All seven hESC lines had similar biological characteristics regardless of karyotype (five normal and two abnormal), such as expression of stage-specific embryonic antigen (SSEA)-4, tumor-rejection antigen (TRA)-1-81 and TRA-1-60 proteins, transcription factor octamer binding protein 4 mRNA, no detectable expression of SSEA-1 protein and high levels of alkaline phosphatase activity. All cell lines were able to undergo differentiation. Imprinted gene expression and DNA methylation were also similar among these cell lines. Non-random X chromosome inactivation patterns were found in XX cell lines.

Conclusions: The present results suggest that hESC lines with abnormal karyotype are also useful experimental materials for cell therapy, developmental biology and genetic research.

Show MeSH
Related in: MedlinePlus