Derivation of human embryonic stem cells in defined conditions

Ludwig, Tenneille E.; Levenstein, Mark E.; Jones, Jeffrey M.; Berggren, W. Travis; Mitchen, Erika R; Frane, Jennifer L.; Crandall, Leann; Daigh, Christine A.; Conard, Kevin R.; Piekarczyk, Marian S.; Llanas, Rachel A.; Thomson, James A.

doi:10.1038/nbt1177

Cited by 1,015 publications

(844 citation statements)

References 15 publications

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“…These cells initially were cultivated on a layer of feeder cells in a serum‐ or KSR‐supplemented medium,104 but in anticipation of clinical applications, the efforts shifted to the development of culture conditions that are feeder‐free and xeno‐free (Table 5). 105, 106, 107, 108, 109, 110, 111 Among these, the E8 medium that was developed by Guokai Chen et al. has become popular.…”

Section: History Of Cell Culture Mediamentioning

confidence: 99%

Animal‐cell culture media: History, characteristics, and current issues

Yao

Asayama

2017

Reprod Medicine & Biology

394

278

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BackgroundCell culture technology has spread prolifically within a century, a variety of culture media has been designed. This review goes through the history, characteristics and current issues of animal‐cell culture media.MethodsA literature search was performed on PubMed and Google Scholar between 1880 and May 2016 using appropriate keywords.ResultsAt the dawn of cell culture technology, the major components of media were naturally derived products such as serum. The field then gradually shifted to the use of chemical‐based synthetic media because naturally derived ingredients have their disadvantages such as large batch‐to‐batch variation. Today, industrially important cells can be cultured in synthetic media. Nevertheless, the combinations and concentrations of the components in these media remain to be optimized. In addition, serum‐containing media are still in general use in the field of basic research. In the fields of assisted reproductive technologies and regenerative medicine, some of the medium components are naturally derived in nearly all instances.ConclusionsFurther improvements of culture media are desirable, which will certainly contribute to a reduction in the experimental variation, enhance productivity among biopharmaceuticals, improve treatment outcomes of assisted reproductive technologies, and facilitate implementation and popularization of regenerative medicine.

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Section: History Of Cell Culture Mediamentioning

confidence: 99%

Animal‐cell culture media: History, characteristics, and current issues

Yao

Asayama

2017

Reprod Medicine & Biology

394

278

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“…However, viral contamination of feeder cells is a common risk in all biotechnological products derived from cell lines, irrespective of the species of origin [22]. For future experiments, we want to carry out the culture of ECSs in a serum-free system in order to eliminate dependence on the culture and the inactivation of murine fibroblasts as well as the risk of contamination with animal pathogens [23].…”

Section: Discussionmentioning

confidence: 99%

Evaluation of a laser technique to isolate the inner cell mass of murine blastocysts

Cortés¹,

Cobo²,

Catalina³

et al. 2007

Biotech and App Biochem

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hESCs (human embryonic stem cells) are pluripotent cells derived from the ICM (inner cell mass) of blastocysts that can be used to derive several kinds of cells of the human body for the treatment of some previously untreated diseases. In considering the future use of hESCs in regenerative medicine and cell-therapy programmes, several research centres have begun projects involving the derivation of hESC lines using spare human embryos from IVF (in vitro fertilization) cycles. In some stem-cell banks, such as ours, the law also permits us to obtain these cell lines. The low availability of spare IVF human embryos, and the low rate of success in the derivation of hESC lines, give these embryos a great research value that limits experiments with new techniques. The use of murine embryos would be a good model with which to do research to discover the best methodologies to use in order to derive new hESC lines. The aim of the present study was to evaluate a new method of isolation of the ICM and derivation of ESC lines in a murine blastocyst model using laser drilling to eliminate the trophectoderm cells and compare it with the usual control method consisting of culturing the whole murine blastocyst. We also tested the adhesion and growth of primary colonies of mESCs (murine ESCs) over two different growth surfaces, namely an MEF (inactive murine fibroblastic feeder layer) or gelatin-coated dishes, in order to achieve the best culture conditions for future derivation of human stem-cell lines for application in human transplantation.

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“…The major constituents of Matrigel are laminin, collagen, and heparan sulphate proteoglycans. Subsequently, various matrices, including laminin (Beattie et al, 2005), fibronectin (Amit et al, 2004) and vitronectin (Braam et al, 2008), and combinations of laminin, fibronectin, and collagen IV with (Ludwig et al, 2006) or without vitronectin (Draper et al, 2004), were reported to support hESC propagation in different media. Most of these ECM proteins are abundant in the human placenta.…”

Section: Hipscs Have Multi-lineage Differentiation Potentialsmentioning

confidence: 99%

“…Nuclei were stained with DAPI (Sigma). FACS analysis was performed as described (Ludwig et al, 2006).…”

Section: Generation Of Hipscsmentioning

confidence: 99%

A novel xeno-free and feeder-cell-free system for human pluripotent stem cell culture

et al. 2012

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While human induced pluripotent stem cells (hiPSCs) have promising applications in regenerative medicine, most of the hiPSC lines available today are not suitable for clinical applications due to contamination with nonhuman materials, such as sialic acid, and potential pathogens from animal-product-containing cell culture systems. Although several xeno-free cell culture systems have been established recently, their use of human fibroblasts as feeders reduces the clinical potential of hiPSCs due to batch-to-batch variation in the feeders and time-consuming preparation processes. In this study, we have developed a xeno-free and feeder-cell-free human embryonic stem cell (hESC)/hiPSC culture system using human plasma and human placenta extracts. The system maintains the self-renewing capacity and pluripotency of hESCs for more than 40 passages. Human iPSCs were also derived from human dermal fibroblasts using this culture system by overexpressing three transcription factors-Oct4, Sox2 and Nanog. The culture system developed here is inexpensive and suitable for large scale production.

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Derivation of human embryonic stem cells in defined conditions

Cited by 1,015 publications

References 15 publications

Animal‐cell culture media: History, characteristics, and current issues

Animal‐cell culture media: History, characteristics, and current issues

Evaluation of a laser technique to isolate the inner cell mass of murine blastocysts

A novel xeno-free and feeder-cell-free system for human pluripotent stem cell culture

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