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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

Derivation of hESC lines and characteristics.(I) Human embryos and isolated ICM. (A) A Day 5 blastocyst cultured in G2.3 blastocyst medium. Note the small ICM. (B) A Day 7 blastocyst that has been cultured in the blastocyst optimum medium for another 2 days. The ICM became clearer. (C) An ICM isolated by mechanical method. (D) A round ICM colony surrounded by a group of residual trophectoderm after mechanical isolation. (E) An ICM isolated by immunosurgery. (F) A dome-like structure on the feeder layer formed after 8 days culture of ICM isolated with immunosurgery. Arrows in C to F indicate ICMs. (G) Typical round hESC colonies with very clear boundary at low magnification. (H) hESC morphology in a colony, showing a high nucleus–cytoplasm ratio; note the presence of nucleoli and typical intercellular spaces at high magnification. Bar = 25 µm in A, B, C and D and bar = 100 µm in E, F and G. (II) Immunocytochemical staining of undifferentiated hESC colonies after 32 passages in four cell lines (A) AP, (B) SSEA-1, (C) SSEA-4, (D) TRA-1-60 and (E) TRA-1-81. Bar = 100 µm.
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DEN137F1: Derivation of hESC lines and characteristics.(I) Human embryos and isolated ICM. (A) A Day 5 blastocyst cultured in G2.3 blastocyst medium. Note the small ICM. (B) A Day 7 blastocyst that has been cultured in the blastocyst optimum medium for another 2 days. The ICM became clearer. (C) An ICM isolated by mechanical method. (D) A round ICM colony surrounded by a group of residual trophectoderm after mechanical isolation. (E) An ICM isolated by immunosurgery. (F) A dome-like structure on the feeder layer formed after 8 days culture of ICM isolated with immunosurgery. Arrows in C to F indicate ICMs. (G) Typical round hESC colonies with very clear boundary at low magnification. (H) hESC morphology in a colony, showing a high nucleus–cytoplasm ratio; note the presence of nucleoli and typical intercellular spaces at high magnification. Bar = 25 µm in A, B, C and D and bar = 100 µm in E, F and G. (II) Immunocytochemical staining of undifferentiated hESC colonies after 32 passages in four cell lines (A) AP, (B) SSEA-1, (C) SSEA-4, (D) TRA-1-60 and (E) TRA-1-81. Bar = 100 µm.

Mentions: In this study, 265 donated embryos were used and 42 (15.8%) developed to early blastocysts on Day 5 (Fig. 1I, A). When these early blastocysts were transferred to blastocyst optimum culture medium for another 2 days, 36 developed to expanded blastocysts and 6 to hatched blastocysts. A total of 42 ICMs (Fig. 1I, B) were isolated using immunosurgery (19 ICM) or mechanical method (23 ICM). All ICMs were seeded on MEF feeder layer (Fig. 1I, C-H).


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)

Derivation of hESC lines and characteristics.(I) Human embryos and isolated ICM. (A) A Day 5 blastocyst cultured in G2.3 blastocyst medium. Note the small ICM. (B) A Day 7 blastocyst that has been cultured in the blastocyst optimum medium for another 2 days. The ICM became clearer. (C) An ICM isolated by mechanical method. (D) A round ICM colony surrounded by a group of residual trophectoderm after mechanical isolation. (E) An ICM isolated by immunosurgery. (F) A dome-like structure on the feeder layer formed after 8 days culture of ICM isolated with immunosurgery. Arrows in C to F indicate ICMs. (G) Typical round hESC colonies with very clear boundary at low magnification. (H) hESC morphology in a colony, showing a high nucleus–cytoplasm ratio; note the presence of nucleoli and typical intercellular spaces at high magnification. Bar = 25 µm in A, B, C and D and bar = 100 µm in E, F and G. (II) Immunocytochemical staining of undifferentiated hESC colonies after 32 passages in four cell lines (A) AP, (B) SSEA-1, (C) SSEA-4, (D) TRA-1-60 and (E) TRA-1-81. Bar = 100 µm.
© Copyright Policy
Related In: Results  -  Collection

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

DEN137F1: Derivation of hESC lines and characteristics.(I) Human embryos and isolated ICM. (A) A Day 5 blastocyst cultured in G2.3 blastocyst medium. Note the small ICM. (B) A Day 7 blastocyst that has been cultured in the blastocyst optimum medium for another 2 days. The ICM became clearer. (C) An ICM isolated by mechanical method. (D) A round ICM colony surrounded by a group of residual trophectoderm after mechanical isolation. (E) An ICM isolated by immunosurgery. (F) A dome-like structure on the feeder layer formed after 8 days culture of ICM isolated with immunosurgery. Arrows in C to F indicate ICMs. (G) Typical round hESC colonies with very clear boundary at low magnification. (H) hESC morphology in a colony, showing a high nucleus–cytoplasm ratio; note the presence of nucleoli and typical intercellular spaces at high magnification. Bar = 25 µm in A, B, C and D and bar = 100 µm in E, F and G. (II) Immunocytochemical staining of undifferentiated hESC colonies after 32 passages in four cell lines (A) AP, (B) SSEA-1, (C) SSEA-4, (D) TRA-1-60 and (E) TRA-1-81. Bar = 100 µm.
Mentions: In this study, 265 donated embryos were used and 42 (15.8%) developed to early blastocysts on Day 5 (Fig. 1I, A). When these early blastocysts were transferred to blastocyst optimum culture medium for another 2 days, 36 developed to expanded blastocysts and 6 to hatched blastocysts. A total of 42 ICMs (Fig. 1I, B) were isolated using immunosurgery (19 ICM) or mechanical method (23 ICM). All ICMs were seeded on MEF feeder layer (Fig. 1I, C-H).

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