Derivation and characterisation of hESC lines from supernumerary embryos, experience from Odense, Denmark

Harkness, Linda; Rasmussen, I.A.; Erb, Karin; Kassem, Moustapha

doi:10.1007/s11626-010-9281-7

Cited by 10 publications

(7 citation statements)

References 22 publications

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“…The derivation, characterization and routine culture of hESC lines (huES9 and KMEB2) has previously been reported [30]–[32]. Cells were cultured on Matrigel (BD Biosciences, San Jose, CA, USA) in mouse embryonic fibroblast-conditioned medium essentially as described in ‘Protocols for Maintenance of Human Embryonic Stem Cells in Feeder Free Conditions’ from Geron Corporation (Menlo Park, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Separate Developmental Programs for HLA-A and -B Cell Surface Expression during Differentiation from Embryonic Stem Cells to Lymphocytes, Adipocytes and Osteoblasts

et al. 2013

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A major problem of allogeneic stem cell therapy is immunologically mediated graft rejection. HLA class I A, B, and Cw antigens are crucial factors, but little is known of their respective expression on stem cells and their progenies. We have recently shown that locus-specific expression (HLA-A, but not -B) is seen on some multipotent stem cells, and this raises the question how this is in other stem cells and how it changes during differentiation. In this study, we have used flow cytometry to investigate the cell surface expression of HLA-A and -B on human embryonic stem cells (hESC), human hematopoietic stem cells (hHSC), human mesenchymal stem cells (hMSC) and their fully-differentiated progenies such as lymphocytes, adipocytes and osteoblasts. hESC showed extremely low levels of HLA-A and no -B. In contrast, multipotent hMSC and hHSC generally expressed higher levels of HLA-A and clearly HLA-B though at lower levels. IFNγ induced HLA-A to very high levels on both hESC and hMSC and HLA-B on hMSC. Even on hESC, a low expression of HLA-B was achieved. Differentiation of hMSC to osteoblasts downregulated HLA-A expression (P = 0.017). Interestingly HLA class I on T lymphocytes differed between different compartments. Mature bone marrow CD4+ and CD8+ T cells expressed similar HLA-A and -B levels as hHSC, while in the peripheral blood they expressed significantly more HLA-B7 (P = 0.0007 and P = 0.004 for CD4+ and CD8+ T cells, respectively). Thus different HLA loci are differentially regulated during differentiation of stem cells.

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Section: Methodsmentioning

confidence: 99%

Separate Developmental Programs for HLA-A and -B Cell Surface Expression during Differentiation from Embryonic Stem Cells to Lymphocytes, Adipocytes and Osteoblasts

et al. 2013

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“…These poor-quality embryos (PQEs) are typically discarded following treatment or donated for research after patient consent. Despite impaired implantation and developmental potential, PQEs have been shown to be a viable source for the derivation of new human embryonic stem cell (hESC) lines [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. It is estimated that hundreds of thousands of embryos per year are discarded because of poor quality and the majority of infertility patients are in favor of donating them for stem cell research [7,17,18].…”

Section: Introductionmentioning

confidence: 99%

“…There have been at least 16 reports describing the feasibility of using PQEs as a source for hESC derivation, with efficiency ranging from 4.1% to 25% [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16]. The variations in hESC derivation efficiency between laboratories are commonly attributed to differences in derivation methodology and experience levels.…”

Section: Introductionmentioning

confidence: 99%

The Influence of Early Embryo Traits on Human Embryonic Stem Cell Derivation Efficiency

O’Leary

Heindryckx

Lierman

et al. 2011

Stem Cells and Development

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Despite its prognostic value in in vitro fertilization, early embryo morphology is not reported on in the derivation of human embryonic stem cell (hESC) lines. Standard hESC derivation does rely on blastocyst development and its efficiency is highly correlated to inner cell mass (ICM) quality. Poor-quality embryos (PQEs) donated for hESC derivation may have a range of cleavage-stage abnormalities that are known to compromise further development. This study was implemented to determine whether specific PQEs traits influence the efficiency of good-quality ICMs to derive new hESC lines. We found that although the types of PQEs investigated were all able to make blastocysts with good-quality ICMs, the ICMs were unequal in their ability to derive hESCs. Good-quality ICMs from embryos with multiple poor-quality traits were unable to generate hESC lines, in contrast to good-quality ICMs from embryos with a single poor-quality trait. In addition, our data suggest a direct correlation between the number of ICM cells present in the blastocyst and its capacity to derive new hESC lines. This study is the first to demonstrate that ICM quality alone is an incomplete indicator of hESC derivation and that application of in vitro fertilization-based early embryo scoring can help predict hESC derivation efficiency. Experiments aiming to quantify, improve upon, or compare hESC derivation efficiency should thus take into consideration early embryo morphology scoring for the comparison of groups with equal developmental competence.

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“…Studies published here did not require ethical approval further to that published in the original derivation articles (Simonsen et al, 2002, Harkness et al, 2010). …”

Section: Methodsmentioning

confidence: 99%

Molecular characterisation of stromal populations derived from human embryonic stem cells: Similarities to immortalised bone marrow derived stromal stem cells

et al. 2015

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Human bone marrow-derived stromal (skeletal) stem cells (BM-hMSC) are being employed in an increasing number of clinical trials for tissue regeneration. A limiting factor for their clinical use is the inability to obtain sufficient cell numbers. Human embryonic stem cells (hESC) can provide an unlimited source of clinical grade cells for therapy. We have generated MSC-like cells from hESC (called here hESC-stromal) that exhibit surface markers and differentiate to osteoblasts and adipocytes, similar to BM-hMSC. In the present study, we used microarray analysis to compare the molecular phenotype of hESC-stromal and immortalised BM-hMSC cells (hMSC-TERT). Of the 7379 genes expressed above baseline, only 9.3% of genes were differentially expressed between undifferentiated hESC-stromal and BM-hMSC. Following ex vivo osteoblast induction, 665 and 695 genes exhibited ≥ 2-fold change (FC) in hESC-stromal and BM-hMSC, respectively with 172 genes common to both cell types. Functional annotation of significantly changing genes revealed similarities in gene ontology between the two cell types. Interestingly, genes in categories of cell adhesion/motility and epithelial–mesenchymal transition (EMT) were highly enriched in hESC-stromal whereas genes associated with cell cycle processes were enriched in hMSC-TERT. This data suggests that while hESC-stromal cells exhibit a similar molecular phenotype to hMSC-TERT, differences exist that can be explained by ontological differences between these two cell types. hESC-stromal cells can thus be considered as a possible alternative candidate cells for hMSC, to be employed in regenerative medicine protocols.

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Derivation and characterisation of hESC lines from supernumerary embryos, experience from Odense, Denmark

Cited by 10 publications

References 22 publications

Separate Developmental Programs for HLA-A and -B Cell Surface Expression during Differentiation from Embryonic Stem Cells to Lymphocytes, Adipocytes and Osteoblasts

Separate Developmental Programs for HLA-A and -B Cell Surface Expression during Differentiation from Embryonic Stem Cells to Lymphocytes, Adipocytes and Osteoblasts

The Influence of Early Embryo Traits on Human Embryonic Stem Cell Derivation Efficiency

Molecular characterisation of stromal populations derived from human embryonic stem cells: Similarities to immortalised bone marrow derived stromal stem cells

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