Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers

Wang, Guangwen; Zhang, Hong; Zhao, Yang; Li, Jian; Cai, Jun; Wang, Peigang; Meng, Sha; Feng, Jing-Bo; Miao, Changxing; Ding, Mei; Li, Dongsheng; Deng, Hongkui

doi:10.1016/j.bbrc.2005.03.058

Cited by 200 publications

(145 citation statements)

References 35 publications

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“…Some require culture on Matrigel (Becton-Dickinson) but this contains a variety of extracellular matrix (ECM) components, most likely associated with an ill-defined mixture of growth factors (20)(21)(22). Others use fully chemically defined media together with specific ECM attachment factors (23).…”

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confidence: 99%

Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium

Furue

Jackson

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

223

207

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A major limitation in developing applications for the use of human embryonic stem cells (HESCs) is our lack of knowledge of their responses to specific cues that control self-renewal, differentiation, and lineage selection. HESCs are most commonly maintained on inactivated mouse embryonic fibroblast feeders in medium supplemented with FCS, or proprietary replacements such as knockout serum-replacement together with FGF-2. These undefined culture conditions hamper analysis of the mechanisms that control HESC behavior. We have now developed a defined serum-free medium, hESF9, for the culture of HESCs on a type I-collagen substrate without feeders. In contrast to other reported media for the culture of HESCs, this medium has a lower osmolarity (292 mosmol/liter), l -ascorbic acid-2-phosphate (0.1 μg/ml), and heparin. Insulin, transferrin, albumin conjugated with oleic acid, and FGF-2 (10 ng/ml) were the only protein components. Further, we found that HESCs would proliferate in the absence of exogenous FGF-2 if heparin was also present. However, their growth was enhanced by the addition of FGF-2 up to 10 ng/ml although higher concentrations were deleterious in the presence of heparin.

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mentioning

confidence: 99%

Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium

Furue

Jackson

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

223

207

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“…Although a number of feeder-free systems have been described (1,3,7,9,10,12,13,21), all hES lines to date have been established and repeatedly passaged on mouse feeders prior to transfer to a feeder free-system for cell propagation. Thus far, the feeder free protocols which we have tried (3,21), have not been compatible with the DA differentiation of hES cell lines developed in this protocol. Finally, it is essential that the bovine serum albumin and porcine transferrin found in KOSR basal media be replaced with human products that can support hES development; currently our efforts (unpublished findings) and those of Gibco (personal communication) in this regard have not been successful.…”

Section: Discussionmentioning

confidence: 99%

“…Currently, there are a number of established hES lines available from NIH-approved sources (Wicell, Bresagen) and a number of new lines provided by the Howard Hughes Institute of Harvard University. All of these cell lines were originally propagated on mouse feeder layers, although several have recently been transferred to feeder free support systems (1,3,7,9,12,21). While the differentiation of DA traits in some hES lines has been described previously (4,5,16,17,18,26), with the exception of one study (19) these protocols involve the prolonged use of complex media containing serum or other undefined reagents and/or cell conditioned media (CM) or co-culture with these cells (ie.…”

Section: Introductionmentioning

confidence: 99%

A protocol for the differentiation of human embryonic stem cells into dopaminergic neurons using only chemically defined human additives: Studies in vitro and in vivo

et al. 2007

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Our ability to use human embryonic stem (hES) cells in cell replacement therapy for Parkinson's disease depends on the discovery of ways to simply and reliably differentiate a dopaminergic (DA) phenotype in these cells. Although several protocols exist for the differentiation of DA traits in hES, they involve the prolonged use of complex media with undefined components, cell conditioned media and/or co-culture with various cells, usually of animal origin. In this study, several well-characterized (H9, BG01) and several new uncharacterized (HUES7, HUES8) hES cell lines were studied for their capacity to differentiate into DA neurons in culture using a novel rapid protocol which uses only chemically-defined human-derived media additives and substrata. Within 3 weeks, cells from all 4 cell lines progressed from the undifferentiated state to β-tubulin III positive cells expressing DA markers in vitro. Moreover, transplantation of these cells into the striata of 6-hydroxydopaminetreated rats at the neuronal progenitor stage resulted in the appearance of differentiated DA traits in vivo 2-3 weeks later.

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“…FGF signaling is crucial for hESC maintenance (20). The addition of bFGF at high concentration (100 ng͞ml) (20,45) or bFGF at low concentration (4 ng͞ml) along with other cytokines, like Noggin (46) and activin͞nodal, can sustain hESC cultures without feeder cells and conditioned medium (47). Wnt3a also can promote hESC proliferation in the absence of a feeder layer or conditioned medium (23).…”

Section: Discussionmentioning

confidence: 99%

Defined culture conditions of human embryonic stem cells

Booth³

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

188

118

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Human embryonic stem cells (hESCs) are pluripotent cells that have the potential to differentiate into any tissue in the human body; therefore, they are a valuable resource for regenerative medicine, drug screening, and developmental studies. However, the clinical application of hESCs is hampered by the difficulties of eliminating animal products in the culture medium and͞or the complexity of conditions required to support hESC growth. We have developed a simple medium [termed hESC Cocktail (HESCO)] containing basic fibroblast growth factor, Wnt3a, April (a proliferation-inducing ligand)͞BAFF (B cell-activating factor belonging to TNF), albumin, cholesterol, insulin, and transferrin, which is sufficient for hESC self-renewal and proliferation. Cells grown in HESCO were maintained in an undifferentiated state as determined by using six different stem cell markers, and their genomic integrity was confirmed by karyotyping. Cells cultured in HESCO readily form embryoid bodies in tissue culture and teratomas in mice. In both cases, the cells differentiated into each of the three cell lineages, ectoderm, endoderm, and mesoderm, indicating that they maintained their pluripotency. The use of a minimal medium sufficient for hESC growth is expected to greatly facilitate clinical application and developmental studies of hESCs.April͞BAFF ͉ fibroblast growth factor ͉ serum free culture ͉ wnt

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Noggin and bFGF cooperate to maintain the pluripotency of human embryonic stem cells in the absence of feeder layers

Cited by 200 publications

References 35 publications

Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium

Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium

A protocol for the differentiation of human embryonic stem cells into dopaminergic neurons using only chemically defined human additives: Studies in vitro and in vivo

Defined culture conditions of human embryonic stem cells

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