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Human embryonic stem cells derived from embryos at different stages of development share similar transcription profiles.

Giritharan G, Ilic D, Gormley M, Krtolica A - PLoS ONE (2011)

Bottom Line: Blastomere-derived hESC lines exhibited all the standard characteristics of hESC including undifferentiated proliferation, genomic stability, expression of pluripotency markers and the ability to differentiate into the cells of all three germ layers both in vitro and in vivo.Furthermore, this profile was evident in very early passages of the cells and did not appear to be affected by extensive passaging.These results suggest that during derivation process cells which give rise to hESC acquire virtually identical stable phenotype and are not affected by the developmental stage of the starting cell population.

View Article: PubMed Central - PubMed

Affiliation: SLL Sciences, StemLifeLine, Inc., San Carlos, California, United States of America.

ABSTRACT
We have derived hESC from biopsied blastomeres of cleavage stage embryos under virtually the same conditions we used for the derivation of hESC lines from inner cell mass of blastocyst stage embryos. Blastomere-derived hESC lines exhibited all the standard characteristics of hESC including undifferentiated proliferation, genomic stability, expression of pluripotency markers and the ability to differentiate into the cells of all three germ layers both in vitro and in vivo. To examine whether hESC lines derived from two developmental stages of the embryo differ in gene expression, we have subjected three blastomere-derived hESC lines and two ICM-derived hESC lines grown under identical culture conditions to transcriptome analysis using gene expression arrays. Unlike previously reported comparisons of hESC lines which demonstrated, apart from core hESC-associated pluripotency signature, significant variations in gene expression profiles of different lines, our data show that hESC lines derived and grown under well-controlled defined culture conditions adopt nearly identical gene expression profiles. Moreover, blastomere-derived and ICM-derived hESC exhibited very similar transcriptional profiles independent of the developmental stage of the embryo from which they originated. Furthermore, this profile was evident in very early passages of the cells and did not appear to be affected by extensive passaging. These results suggest that during derivation process cells which give rise to hESC acquire virtually identical stable phenotype and are not affected by the developmental stage of the starting cell population.

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Differentially expressed genes with fold change after direct comparison of cleavage stage blastomere-derived (SBD) and blastocyst stage whole embryo-derived (WED) lines.Blue and red colors represent lower and higher expression, respectively.
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pone-0026570-g005: Differentially expressed genes with fold change after direct comparison of cleavage stage blastomere-derived (SBD) and blastocyst stage whole embryo-derived (WED) lines.Blue and red colors represent lower and higher expression, respectively.

Mentions: To identify the genes which show significant differences between blastomere and ICM-derived lines, direct pair-wise comparison was performed. The analysis revealed only 36 annotated genes (20 genes upregulated in blastomere derived lines relative to ICM-derived and 16 genes downregulated) were significantly different between the lines of both origins (Fig. 5). Out of these 36 genes, only 6 genes (3 upregulated - ZNF248, SOHLH2 and MAGEE1, and 3 downregulated - CLC, ZNF558 and LGALS14) showed more than 4 fold changes in expression between these hESC lines indicating that only minor differences exist. These include three DNA binding proteins with potential role in regulation of gene transcription: ZNF248, ZNF558 and SOHLH2 and proteins with potential membrane functions: MAGEE1, LGALS14 and CLC (Charcot-Leyden crystal protein/Galectn-10). ZNF248 is a Krueppel C2H2-type zinc-finger DNA binding protein with potential role in transcriptional regulation [16]. ZNF558 is zinc finger DNA binding protein that also binds to Rrp46 which is an exosome subunit and mRNA splicing/processing factor with potential role in mRNA degradation and gene expression [17]. SOHLH2 is a spermatogenesis- and oogenesis-specific basic helix–loop–helix (bHLH) transcription factor implicated in regulation of early germ cell development [18]. MAGEE1 gene is an X chromosome gene and encodes an α-dystrobrevin-associated MAGE (melanoma-associated antigen) protein (DAMAGE) that may have a signaling role in brain, muscle, and peripheral nerve [19]. LGALS14 gene encodes a galectin family protein predominantly expressed in placenta [20]. CLC gene encodes lysophospholipase, enzymes that regulate the lysophospholipids in biological membranes [21]. It is related to the galectin family and may also possess carbohydrate or IgE-binding activities, and be associated with inflammation and some myeloid leukemias [22].


Human embryonic stem cells derived from embryos at different stages of development share similar transcription profiles.

Giritharan G, Ilic D, Gormley M, Krtolica A - PLoS ONE (2011)

Differentially expressed genes with fold change after direct comparison of cleavage stage blastomere-derived (SBD) and blastocyst stage whole embryo-derived (WED) lines.Blue and red colors represent lower and higher expression, respectively.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198782&req=5

pone-0026570-g005: Differentially expressed genes with fold change after direct comparison of cleavage stage blastomere-derived (SBD) and blastocyst stage whole embryo-derived (WED) lines.Blue and red colors represent lower and higher expression, respectively.
Mentions: To identify the genes which show significant differences between blastomere and ICM-derived lines, direct pair-wise comparison was performed. The analysis revealed only 36 annotated genes (20 genes upregulated in blastomere derived lines relative to ICM-derived and 16 genes downregulated) were significantly different between the lines of both origins (Fig. 5). Out of these 36 genes, only 6 genes (3 upregulated - ZNF248, SOHLH2 and MAGEE1, and 3 downregulated - CLC, ZNF558 and LGALS14) showed more than 4 fold changes in expression between these hESC lines indicating that only minor differences exist. These include three DNA binding proteins with potential role in regulation of gene transcription: ZNF248, ZNF558 and SOHLH2 and proteins with potential membrane functions: MAGEE1, LGALS14 and CLC (Charcot-Leyden crystal protein/Galectn-10). ZNF248 is a Krueppel C2H2-type zinc-finger DNA binding protein with potential role in transcriptional regulation [16]. ZNF558 is zinc finger DNA binding protein that also binds to Rrp46 which is an exosome subunit and mRNA splicing/processing factor with potential role in mRNA degradation and gene expression [17]. SOHLH2 is a spermatogenesis- and oogenesis-specific basic helix–loop–helix (bHLH) transcription factor implicated in regulation of early germ cell development [18]. MAGEE1 gene is an X chromosome gene and encodes an α-dystrobrevin-associated MAGE (melanoma-associated antigen) protein (DAMAGE) that may have a signaling role in brain, muscle, and peripheral nerve [19]. LGALS14 gene encodes a galectin family protein predominantly expressed in placenta [20]. CLC gene encodes lysophospholipase, enzymes that regulate the lysophospholipids in biological membranes [21]. It is related to the galectin family and may also possess carbohydrate or IgE-binding activities, and be associated with inflammation and some myeloid leukemias [22].

Bottom Line: Blastomere-derived hESC lines exhibited all the standard characteristics of hESC including undifferentiated proliferation, genomic stability, expression of pluripotency markers and the ability to differentiate into the cells of all three germ layers both in vitro and in vivo.Furthermore, this profile was evident in very early passages of the cells and did not appear to be affected by extensive passaging.These results suggest that during derivation process cells which give rise to hESC acquire virtually identical stable phenotype and are not affected by the developmental stage of the starting cell population.

View Article: PubMed Central - PubMed

Affiliation: SLL Sciences, StemLifeLine, Inc., San Carlos, California, United States of America.

ABSTRACT
We have derived hESC from biopsied blastomeres of cleavage stage embryos under virtually the same conditions we used for the derivation of hESC lines from inner cell mass of blastocyst stage embryos. Blastomere-derived hESC lines exhibited all the standard characteristics of hESC including undifferentiated proliferation, genomic stability, expression of pluripotency markers and the ability to differentiate into the cells of all three germ layers both in vitro and in vivo. To examine whether hESC lines derived from two developmental stages of the embryo differ in gene expression, we have subjected three blastomere-derived hESC lines and two ICM-derived hESC lines grown under identical culture conditions to transcriptome analysis using gene expression arrays. Unlike previously reported comparisons of hESC lines which demonstrated, apart from core hESC-associated pluripotency signature, significant variations in gene expression profiles of different lines, our data show that hESC lines derived and grown under well-controlled defined culture conditions adopt nearly identical gene expression profiles. Moreover, blastomere-derived and ICM-derived hESC exhibited very similar transcriptional profiles independent of the developmental stage of the embryo from which they originated. Furthermore, this profile was evident in very early passages of the cells and did not appear to be affected by extensive passaging. These results suggest that during derivation process cells which give rise to hESC acquire virtually identical stable phenotype and are not affected by the developmental stage of the starting cell population.

Show MeSH
Related in: MedlinePlus