A Survey of Parameters Involved in the Establishment of New Lines of Human Embryonic Stem Cells

Fraga, Ana; Araújo, Érica Sara Souza de; Stabellini, Raquel; Vergani, Naja; Pereira, Lygia V.

doi:10.1007/s12015-011-9250-x

Cited by 21 publications

(23 citation statements)

References 41 publications

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“…LIF is one of the genes expressed in unrestricted somatic stem cells, which have been described as recapitulating the characteristics of mesenchymal stem cells (MSCs) from placental cord blood and, among the recently proposed transcriptome-based subtypes of MSCs [23], are therefore the most closely related to human embryonic stem cells (hESCs). Accordingly, LIF (alone or in combination with bFGF), is the main factor maintaining the pluripotency of hESCs during derivation and long-term culture [36]. It has more recently been demonstrated that LIF induces the expression of miRNA-21, a miRNA that promotes EMT through STAT3 signalling in human tumor cells [19] and was shown in our ISH experiments to be expressed at similar levels in all three classes.…”

Section: Discussionmentioning

confidence: 62%

New transcriptional-based insights into the pathogenesis of desmoplastic small round cell tumors (DSRCTs)

et al. 2017

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To gain new insights into desmoplastic small round cell tumors (DSRCTs) by means of gene expression profiling (GEP). Formalin-fixed, paraffin-embedded surgical specimens obtained from seven pretreated DSRCT patients were interrogated using GEP complemented by immunohistochemistry, a cancer stem cell array, and miRNA in situ hybridisation, including the combined chimera modules miRNA-200/ZEB1 and miRNA-34/SLUG. The chimera modules divided the cases into three classes that respectively recapitulated the traits of mesenchymal epithelial reverse transition (MErT), epithelial mesenchymal transition (EMT), and hybrid/partial EMT. This indicates a close correlation between the reprogramming governed by EMT regulators and DSRCT biology, which was further confirmed by miRNA-21 and is consistent with the broad morphological spectrum of DSRCTs. Starting from the miRNA-200/ZEB1 axis, we also found that DSRCTs carry a signature of immunological ignorance that is not responsive to PD-L1 blockade. Evidence that the up-regulation of miRNA-200 and E-cadherin, and quite a high level of miRNA-21 expression segregate with the MErT supports the idea that, in addition to the hybrid/partial state, MErT is also enriched in stemness: the androgen-positive cases, whose stemness traits were confirmed by stem cell arrays, all fell into these two classes. Our findings also confirmed that tumoral cell PDGFRA expression correlates with desmoplasia, and demonstrated the co-expression of PDGFRA and ISLR/Meflin, another marker of pluripotency. Despite the limited number of cases, these findings provide unexpectedly relevant information concerning the pathogenesis of DSRCTs, and prove the validity of miRNA-based chimera circuit modelling in the clinico-pathological setting.

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Section: Discussionmentioning

confidence: 62%

New transcriptional-based insights into the pathogenesis of desmoplastic small round cell tumors (DSRCTs)

et al. 2017

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“…7 To date, more than 1000 hES cell lines have been derived in laboratories worldwide, and rapid advances in the knowledge and technologies associated with the culture conditions of hES cells have made it possible to derive new hES cell lines under clinical or near clinical conditions for further use. 8 Although the embryos may display poor morphology, they remain a major source of starting material for the isolation of hES cells, and several methods have been applied to improve the success of hES cell derivation. Culturing poor-quality embryos in a modified culture medium increased blastocyst formation, 9 and using mesenchymal stem cells as feeder cells also facilitated the derivation of hES cells from poor-quality embryos.…”

Section: Introductionmentioning

confidence: 99%

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

Pruksananonda

Rungsiwiwut

Numchaisrika

et al. 2012

BioResearch Open Access

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Human embryonic stem (hES) cells are considered to be a potential source for the therapy of human diseases, drug screening, and the study of developmental biology. In the present study, we successfully derived hES cell lines from blastocysts developed from frozen and fresh embryos. Seventeen- to eighteen-year-old frozen embryos were thawed, cultured to the blastocyst stage, and induced to form hES cells using human foreskin fibroblasts. The Chula2.hES cell line and the Chula4.hES and Chula5.hES cell lines were derived from blastocysts developed from frozen and fresh embryos, respectively. The cell lines expressed pluripotent markers, including alkaline phosphatase (AP), Oct3/4, stage-specific embryonic antigen (SSEA)-4, and tumor recognition antigen (TRA)-1-60 and TRA-1-81 as detected with immunocytochemistry. The real-time polymerase chain reaction (RT-PCR) results showed that the cell lines expressed pluripotent genes, including OCT3/4, SOX2, NANOG, UTF, LIN28, REX1, NODAL, and E-Cadherin. In addition, the telomerase activities of the cell lines were higher than in the fibroblast cells. Moreover, the cell lines differentiated into all three germ layers both in vitro and in vivo. The cell lines had distinct identities, as revealed with DNA fingerprinting, and maintained their normal karyotype after a long-term culture. This study is the first to report the successful derivation of hES cell lines in Thailand and that frozen embryos maintained their pluripotency similar to fresh embryos, as shown by the success of hES cell derivation, even after years of cryopreservation. Therefore, embryos from prolonged cryopreservation could be an alternative source for embryonic stem cell research.

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“…The first in-man transfusion is likely to be from one of these two cell sources with studies of red cells generated from hESC or hiPSC coming later [170–172]. Currently 1200 hESC lines have been established worldwide, and 375 of these are deposited in two international registries [172].…”

Section: Resultsmentioning

confidence: 99%

“…Currently 1200 hESC lines have been established worldwide, and 375 of these are deposited in two international registries [172]. Investigators at the forefront of ex vivo expansion studies expect that GMP compliant facilities to produce red cells ex vivo will be available within the next 2-3 years.…”

Section: Resultsmentioning

confidence: 99%

“…Theoretical calculations to determine how many iPSC lines would be needed to support alloimmunized patients in France have already been performed [171]. Although a large number of embryonic stem cell lines have been developed, the expression of blood group antigens by erythrocytes from these cell lines is not known [172]. Forty percent of rare units in France were provided to patients with sickle cell anemia [171].…”

Section: Resultsmentioning

confidence: 99%

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Alternative Blood Products and Clinical Needs in Transfusion Medicine

Whitsett

Vaglio

Grazzini

2012

Stem Cells International

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The primary focus of national blood programs is the provision of a safe and adequate blood supply. This goal is dependent on regular voluntary donations and a regulatory infrastructure that establishes and enforces standards for blood safety. Progress in ex vivo expansion of blood cells from cell sources including peripheral blood, cord blood, induced pluripotent stem cells, and human embryonic stem cell lines will likely make alternative transfusion products available for clinical use in the near future. Initially, alloimmunized patients and individuals with rare blood types are most likely to benefit from alternative products. However, in developed nations voluntary blood donations are projected to be inadequate in the future as blood usage by individuals 60 years and older increases. In developing nations economic and political challenges may impede progress in attaining self-sufficiency. Under these circumstances, ex vivo generated red cells may be needed to supplement the general blood supply.

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A Survey of Parameters Involved in the Establishment of New Lines of Human Embryonic Stem Cells

Cited by 21 publications

References 41 publications

New transcriptional-based insights into the pathogenesis of desmoplastic small round cell tumors (DSRCTs)

New transcriptional-based insights into the pathogenesis of desmoplastic small round cell tumors (DSRCTs)

Eighteen-Year Cryopreservation Does Not Negatively Affect the Pluripotency of Human Embryos: Evidence from Embryonic Stem Cell Derivation

Alternative Blood Products and Clinical Needs in Transfusion Medicine

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