Mechanical isolation of the inner cell mass is effective in derivation of new human embryonic stem cell lines

Ström, Susanne; Inzunza, José; Grinnemo, Karl‐Henrik; Holmberg, K.; Matilainen, Eija; Strömberg, Anne‐Marie; Blennow, Elisabeth; Hovatta, Outi

doi:10.1093/humrep/dem335

Cited by 94 publications

(66 citation statements)

References 23 publications

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“…Efficient methods to derive and grow hESCs remain challenging even as methods are optimized for their derivation from embryos of variable quality (Ström et al, 2007). Early steps in the derivation process are crucial, as the majority of embryos either fail to generate a viable ICM or the isolated ICM cells differentiate upon initial passaging.…”

Section: Deriving and Culturing Hescs From Pre-implantation Embryosmentioning

confidence: 99%

“…Early steps in the derivation process are crucial, as the majority of embryos either fail to generate a viable ICM or the isolated ICM cells differentiate upon initial passaging. Although hESCs have been established from plating whole blastocysts, the derivation of hESCs more typically involves immunosurgery, microdissection or laser-mediated separation and destruction of the TE prior to culturing the ICM in vitro (Thomson et al, 1998;Ström et al, 2007; Chen et al, 2009). The benefit of TE elimination during hESC derivation is incompletely understood and might be due to the removal of negative, possibly paracrine, signals originating from the TE that cause the differentiation of ICM-derived cells in vitro.…”

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confidence: 99%

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Human pre-implantation embryo development

et al. 2012

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SummaryUnderstanding human pre-implantation development has important implications for assisted reproductive technology (ART) and for human embryonic stem cell (hESC)-based therapies. Owing to limited resources, the cellular and molecular mechanisms governing this early stage of human development are poorly understood. Nonetheless, recent advances in non-invasive imaging techniques and molecular and genomic technologies have helped to increase our understanding of this fascinating stage of human development. Here, we summarize what is currently known about human pre-implantation embryo development and highlight how further studies of human pre-implantation embryos can be used to improve ART and to fully harness the potential of hESCs for therapeutic goals. Key words: Human embryo, Oocyte to embryo transition, Zygote, Cleavage division, Blastocyst, Human embryonic stem cells, Aneuploidy IntroductionStudies of mammalian embryo development, especially in the mouse, have provided key insights into early mammalian developmental pathways. However, species-specific differences, for example in the timing of the major wave of genome activation, the patterns of gene expression, the frequency of chromosome missegregation and the patterns of epigenetic modifications, may limit the extrapolation of some findings to human embryo development. To date, studies of human pre-implantation development have utilized spare human pre-implantation embryos derived from in vitro fertilization (IVF; see Glossary, Box 1) to obtain insights into aspects of development specific to humans. Historically, these studies have focused on the morphological examination of embryos and the identification of factors that can improve in vitro culture, such as the conditions required to fertilize human oocytes in vitro, to cryopreserve and thaw human embryos and to promote human blastocyst formation. More recently, with the advent of advanced imaging techniques and sensitive gene expression profiling technologies, these studies are beginning to provide a clearer understanding of human pre-implantation development at cellular and molecular resolution. In addition, spare human pre-implantation embryos have enabled the derivation of human embryonic stem cells (hESCs), leading to the establishment of novel tools for human developmental biology and the emergence of a new field of research, namely hESC-based regenerative medicine. Thus, further investigations into the fundamental aspects of human pre-implantation development might provide not only insights into human developmental biology and common birth defects but also potential benefits for reproductive health and improvements in regenerative medicine.In this Primer, we delve further into the specifics of human preimplantation embryo development and discuss its relationship to that of other species. We also provide a summary of our current understanding of the molecular pathways of early human embryo development. Finally, we discuss how studies of hESCs can be used to further our understanding of early hum...

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Section: Deriving and Culturing Hescs From Pre-implantation Embryosmentioning

confidence: 99%

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confidence: 99%

Human pre-implantation embryo development

et al. 2012

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“…growth factors, signaling pathways and oxygen pressure Powers et al 2008). Pluripotency of embryonic stem cells is often concluded with the formation of in vivo teratoma, which contains cells from the three germ layers, when injecting clusters of embryonic stem cells in SCID mice (Prokhorova et al 2008;Strom et al 2007;Valbuena et al 2006). The drawbacks of this technique is that in vivo teratoma does not show the differentiation potential of one single hESC colony clone as several hESC clusters are injected into the host animal.…”

Section: Discussionmentioning

confidence: 99%

Sustained embryoid body formation and culture in a non-laborious three dimensional culture system for human embryonic stem cells

Stenberg

Elovsson

Strehl³

et al. 2011

Cytotechnology

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Pluripotent human embryonic stem cell (hESC) lines are a promising model system in developmental and tissue regeneration research. Differentiation of hESCs towards the three germ layers and finally tissue specific cell types is often performed through the formation of embryoid bodies (EBs) in suspension or hanging droplet culture systems. However, these systems are inefficient regarding embryoid body (EB) formation, structural support to the EB and long term differentiation capacity. The present study investigates if agarose, as a semi solid matrix, can facilitate EB formation and support differentiation of hESC lines. The results showed that agarose culture is able to enhance EB formation efficiency with 10% and increase EB growth by 300%. The agarose culture system was able to maintain expression of the three germ layers over 8 weeks of culture. All of the four hESC lines tested developed EBs in the agarose system although with a histological heterogeneity between cell lines as well as within cell lines. In conclusion, a 3-D agarose culture of spherical hESC colonies improves EB formation and growth in a cost effective, stable and non-laborious technique.

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“…Their study indicated that this approach is potentially useful in xenofree hESC derivation because it avoids the use of animal-derived enzymes of immunological substances that have traditionally been used for this purpose. Ström et al developed another potentially advantageous isolation method wherein a mechanical isolation protocol utilizes two metal needles with sharpened tips that can cut both the zona pellucida and ICM, thereby avoiding exposure of embryos to either acidic environments or animal-derived substances (Strom et al 2007 ) .…”

Section: Studies Involved In Improving Isolation and Cell Culture Conmentioning

confidence: 99%

Mesenchymal Stem Cells: Possibilities of New Treatment Options

Tokcaer-Keskin

Koçak

Gürsel

et al. 2012

Adult and Embryonic Stem Cells

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Mechanical isolation of the inner cell mass is effective in derivation of new human embryonic stem cell lines

Abstract: Mechanical isolation of the ICM proved to be an effective way to derive new hESC lines. The technique is fast, does not require any extra investment and the xeno-components of immunosurgery could be avoided.

Cited by 94 publications

References 23 publications

Human pre-implantation embryo development

Human pre-implantation embryo development

Sustained embryoid body formation and culture in a non-laborious three dimensional culture system for human embryonic stem cells

Mesenchymal Stem Cells: Possibilities of New Treatment Options

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