Limits...
Long-term expandable SOX9+ chondrogenic ectomesenchymal cells from human pluripotent stem cells.

Umeda K, Oda H, Yan Q, Matthias N, Zhao J, Davis BR, Nakayama N - Stem Cell Reports (2015)

Bottom Line: When "primed" with TGFβ, such cells efficiently formed translucent cartilage particles, which were completely mineralized in 12 weeks in immunocompromized mice.Ectomesenchyme is a source of many craniofacial bone and cartilage structures.The method we describe for obtaining a large quantity of human ectomesenchymal cells will help to model craniofacial disorders in vitro and potentially provide cells for the repair of craniofacial damage.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Medicine, The University of Texas Health Science Center at Houston Medical School, Houston, TX 77030, USA.

Show MeSH

Related in: MedlinePlus

Comparative Transcriptome Analysis of Ectomesenchymal Cells Using the RNA-seq Technology(A) Heat map of the top 250 genes, which are differentially expressed among the three groups. Lanes 1–3: “FSB”; lanes 4, 5, 8: “F”; lanes 6, 7, 9: “FSB→F.”(B) Principal component analysis of the expression pattern of protein-coding genes among the three groups. Blue, “FSB”; red, “F”; green, “FSB→F.”(C) Selected list of culture condition-specific genes and common genes. Genes that gave a p < 0.05 were picked from Tables S3 and S4 except for few with ∗ that gave slightly more variations (0.05 < p < 0.1). ND, not determined.(D) GO analysis. Selected GO categories from Table S1 (upper) and a selected list of development-related GO terms from Table S2 (lower).(E) RT-PCR confirmation of the relative gene expression levels predicted by the FPKM values (n = 3 independent cultures, mean ± SD).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4400647&req=5

fig6: Comparative Transcriptome Analysis of Ectomesenchymal Cells Using the RNA-seq Technology(A) Heat map of the top 250 genes, which are differentially expressed among the three groups. Lanes 1–3: “FSB”; lanes 4, 5, 8: “F”; lanes 6, 7, 9: “FSB→F.”(B) Principal component analysis of the expression pattern of protein-coding genes among the three groups. Blue, “FSB”; red, “F”; green, “FSB→F.”(C) Selected list of culture condition-specific genes and common genes. Genes that gave a p < 0.05 were picked from Tables S3 and S4 except for few with ∗ that gave slightly more variations (0.05 < p < 0.1). ND, not determined.(D) GO analysis. Selected GO categories from Table S1 (upper) and a selected list of development-related GO terms from Table S2 (lower).(E) RT-PCR confirmation of the relative gene expression levels predicted by the FPKM values (n = 3 independent cultures, mean ± SD).

Mentions: We performed genome-wide, comparative transcriptome analyses to elucidate the molecular basis of the effect of Nodal/Activin/TGFβ signaling-suppression on the maintenance of the proliferative and chondrogenic properties of the CD271+CD73+ ectomesenchymal cells. The CD271hiCD73− neural crest-like progeny of H9 hESCs were isolated and expanded under FGF2+SB431542 conditions for six to seven passages (FSB, Figure 6). In some cases, the culture condition was shifted to the FGF2 condition from p4 (FSB→F). As a control, the CD271hiCD73− cells were cultured from the outset under FGF2-conditions (F) for six passages.


Long-term expandable SOX9+ chondrogenic ectomesenchymal cells from human pluripotent stem cells.

Umeda K, Oda H, Yan Q, Matthias N, Zhao J, Davis BR, Nakayama N - Stem Cell Reports (2015)

Comparative Transcriptome Analysis of Ectomesenchymal Cells Using the RNA-seq Technology(A) Heat map of the top 250 genes, which are differentially expressed among the three groups. Lanes 1–3: “FSB”; lanes 4, 5, 8: “F”; lanes 6, 7, 9: “FSB→F.”(B) Principal component analysis of the expression pattern of protein-coding genes among the three groups. Blue, “FSB”; red, “F”; green, “FSB→F.”(C) Selected list of culture condition-specific genes and common genes. Genes that gave a p < 0.05 were picked from Tables S3 and S4 except for few with ∗ that gave slightly more variations (0.05 < p < 0.1). ND, not determined.(D) GO analysis. Selected GO categories from Table S1 (upper) and a selected list of development-related GO terms from Table S2 (lower).(E) RT-PCR confirmation of the relative gene expression levels predicted by the FPKM values (n = 3 independent cultures, mean ± SD).
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4400647&req=5

fig6: Comparative Transcriptome Analysis of Ectomesenchymal Cells Using the RNA-seq Technology(A) Heat map of the top 250 genes, which are differentially expressed among the three groups. Lanes 1–3: “FSB”; lanes 4, 5, 8: “F”; lanes 6, 7, 9: “FSB→F.”(B) Principal component analysis of the expression pattern of protein-coding genes among the three groups. Blue, “FSB”; red, “F”; green, “FSB→F.”(C) Selected list of culture condition-specific genes and common genes. Genes that gave a p < 0.05 were picked from Tables S3 and S4 except for few with ∗ that gave slightly more variations (0.05 < p < 0.1). ND, not determined.(D) GO analysis. Selected GO categories from Table S1 (upper) and a selected list of development-related GO terms from Table S2 (lower).(E) RT-PCR confirmation of the relative gene expression levels predicted by the FPKM values (n = 3 independent cultures, mean ± SD).
Mentions: We performed genome-wide, comparative transcriptome analyses to elucidate the molecular basis of the effect of Nodal/Activin/TGFβ signaling-suppression on the maintenance of the proliferative and chondrogenic properties of the CD271+CD73+ ectomesenchymal cells. The CD271hiCD73− neural crest-like progeny of H9 hESCs were isolated and expanded under FGF2+SB431542 conditions for six to seven passages (FSB, Figure 6). In some cases, the culture condition was shifted to the FGF2 condition from p4 (FSB→F). As a control, the CD271hiCD73− cells were cultured from the outset under FGF2-conditions (F) for six passages.

Bottom Line: When "primed" with TGFβ, such cells efficiently formed translucent cartilage particles, which were completely mineralized in 12 weeks in immunocompromized mice.Ectomesenchyme is a source of many craniofacial bone and cartilage structures.The method we describe for obtaining a large quantity of human ectomesenchymal cells will help to model craniofacial disorders in vitro and potentially provide cells for the repair of craniofacial damage.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecular Medicine, The University of Texas Health Science Center at Houston Medical School, Houston, TX 77030, USA.

Show MeSH
Related in: MedlinePlus