Millipore Sigma Vibrant Logo
 

neural+stem+cell+expansion+media


15 Results Advanced Search  
Showing
Products (1)
Documents (5)
Site Content (9)
Can't Find What You're Looking For?
Contact Customer Service

 
  • «
  • <
  • 1
  • >
  • »

  • Process-based expansion and neural differentiation of human pluripotent stem cells for transplantation and disease modeling. 23893392

    Robust strategies for developing patient-specific, human, induced pluripotent stem cell (iPSC)-based therapies of the brain require an ability to derive large numbers of highly defined neural cells. Recent progress in iPSC culture techniques includes partial-to-complete elimination of feeder layers, use of defined media, and single-cell passaging. However, these techniques still require embryoid body formation or coculture for differentiation into neural stem cells (NSCs). In addition, none of the published methodologies has employed all of the advances in a single culture system. Here we describe a reliable method for long-term, single-cell passaging of PSCs using a feeder-free, defined culture system that produces confluent, adherent PSCs that can be differentiated into NSCs. To provide a basis for robust quality control, we have devised a system of cellular nomenclature that describes an accurate genotype and phenotype of the cells at specific stages in the process. We demonstrate that this protocol allows for the efficient, large-scale, cGMP-compliant production of transplantable NSCs from all lines tested. We also show that NSCs generated from iPSCs produced with the process described are capable of forming both glia defined by their expression of S100β and neurons that fire repetitive action potentials.
    Document Type:
    Reference
    Product Catalog Number:
    Multiple
    Product Catalog Name:
    Multiple

  • Optimization and integration of expansion and neural commitment of mouse embryonic stem cells. 17596123

    To harness the potential of ES (embryonic stem) cells for human therapy, technology to develop the large-scale expansion and differentiation of these cells is required. In the present study, we tested various conditions for the expansion and neural commitment of mouse ES cells, using a cell line with a fluorescent reporter, which allows the monitoring of these processes by flow cytometry. The expansion of the 46C ES cell line in the presence of two different media [serum-free ESGRO Completetrade mark and DMEM (Dulbecco's modified Eagle's medium) containing 10% (v/v) fetal bovine serum] was compared. Both media yielded similar cell fold increases at two different initial cell densities and were able to maintain neural commitment potential during expansion. The influence of inocula concentration in the presence of two different media on cell proliferation and efficiency of neural commitment was evaluated. Two different chemically defined serum-free media were tested: the more conventional N2B27 and the second-generation medium RHB-A (Stem Cell Sciences, Edinburgh, Scotland, U.K.). The kinetics of neural commitment was followed during 8 days in the presence of both media. Our results show that inocula concentrations between 5x10(3) and 10(4) cells/cm(2) are the most appropriate to achieve a better cell growth and more efficient neural commitment. We also show that cell culture in RHB-A medium results in higher rates of cell proliferation and neural commitment of ES cells, when compared with N2B27.
    Document Type:
    Reference
    Product Catalog Number:
    SCM012
    Product Catalog Name:
    NDiff Neuro-2 Medium Supplement (200x)

  • Neurotrophic effects of leukemia inhibitory factor on neural cells derived from human embryonic stem cells. 22899336

    Various growth factor cocktails have been used to proliferate and then differentiate human neural progenitor (NP) cells derived from embryonic stem cells (ESC) for in vitro and in vivo studies. However, the cytokine leukemia inhibitory factor (LIF) has been largely overlooked. Here, we demonstrate that LIF significantly enhanced in vitro survival and promoted differentiation of human ESC-derived NP cells. In NP cells, as well as NP-derived neurons, LIF reduced caspase-mediated apoptosis and reduced both spontaneous and H(2) O(2) -induced reactive oxygen species in culture. In vitro, NP cell proliferation and the yield of differentiated neurons were significantly higher in the presence of LIF. In NP cells, LIF enhanced cMyc phosphorylation, commonly associated with self-renewal/proliferation. Also, in differentiating NP cells LIF activated the phosphoinositide 3-kinase and signal transducer and activator of transcription 3 pathways, associated with cell survival and reduced apoptosis. When differentiated in LIF(+) media, neurite outgrowth and ERK1/2 phosphorylation were potentiated together with increased expression of gp130, a component of the LIF receptor complex. NP cells, pretreated in vitro with LIF, were effective in reducing infarct volume in a model of focal ischemic stroke but LIF did not lead to significantly improved initial NP cell survival over nontreated NP cells. Our results show that LIF signaling significantly promotes human NP cell proliferation, survival, and differentiation in vitro. Activated LIF signaling should be considered in cell culture expansion systems for future human NP cell-based therapeutic transplant studies. STEM CELLS2012;30:2387-2399.
    Document Type:
    Reference
    Product Catalog Number:
    AB5622
    Product Catalog Name:
    Anti-Microtubule-Associated Protein 2 (MAP2) Antibody

  • Niche-independent symmetrical self-renewal of a mammalian tissue stem cell. 16086633

    Pluripotent mouse embryonic stem (ES) cells multiply in simple monoculture by symmetrical divisions. In vivo, however, stem cells are generally thought to depend on specialised cellular microenvironments and to undergo predominantly asymmetric divisions. Ex vivo expansion of pure populations of tissue stem cells has proven elusive. Neural progenitor cells are propagated in combination with differentiating progeny in floating clusters called neurospheres. The proportion of stem cells in neurospheres is low, however, and they cannot be directly observed or interrogated. Here we demonstrate that the complex neurosphere environment is dispensable for stem cell maintenance, and that the combination of fibroblast growth factor 2 (FGF-2) and epidermal growth factor (EGF) is sufficient for derivation and continuous expansion by symmetrical division of pure cultures of neural stem (NS) cells. NS cells were derived first from mouse ES cells. Neural lineage induction was followed by growth factor addition in basal culture media. In the presence of only EGF and FGF-2, resulting NS cells proliferate continuously, are diploid, and clonogenic. After prolonged expansion, they remain able to differentiate efficiently into neurons and astrocytes in vitro and upon transplantation into the adult brain. Colonies generated from single NS cells all produce neurons upon growth factor withdrawal. NS cells uniformly express morphological, cell biological, and molecular features of radial glia, developmental precursors of neurons and glia. Consistent with this profile, adherent NS cell lines can readily be established from foetal mouse brain. Similar NS cells can be generated from human ES cells and human foetal brain. The extrinsic factors EGF plus FGF-2 are sufficient to sustain pure symmetrical self-renewing divisions of NS cells. The resultant cultures constitute the first known example of tissue-specific stem cells that can be propagated without accompanying differentiation. These homogenous cultures will enable delineation of molecular mechanisms that define a tissue-specific stem cell and allow direct comparison with pluripotent ES cells.
    Document Type:
    Reference
    Product Catalog Number:
    SCM012
    Product Catalog Name:
    NDiff Neuro-2 Medium Supplement (200x)

  • A defined and xeno-free culture method enabling the establishment of clinical-grade human embryonic, induced pluripotent and adipose stem cells. 20419109

    The growth of stem cells in in vitro conditions requires optimal balance between signals mediating cell survival, proliferation, and self-renewal. For clinical application of stem cells, the use of completely defined conditions and elimination of all animal-derived materials from the establishment, culture, and differentiation processes is desirable.Here, we report the development of a fully defined xeno-free medium (RegES), capable of supporting the expansion of human embryonic stem cells (hESC), induced pluripotent stem cells (iPSC) and adipose stem cells (ASC). We describe the use of the xeno-free medium in the derivation and long-term (greater than 80 passages) culture of three pluripotent karyotypically normal hESC lines: Regea 06/015, Regea 07/046, and Regea 08/013. Cardiomyocytes and neural cells differentiated from these cells exhibit features characteristic to these cell types. The same formulation of the xeno-free medium is capable of supporting the undifferentiated growth of iPSCs on human feeder cells. The characteristics of the pluripotent hESC and iPSC lines are comparable to lines derived and cultured in standard undefined culture conditions. In the culture of ASCs, the xeno-free medium provided significantly higher proliferation rates than ASCs cultured in medium containing allogeneic human serum (HS), while maintaining the differentiation potential and characteristic surface marker expression profile of ASCs, although significant differences in the surface marker expression of ASCs cultured in HS and RegES media were revealed.Our results demonstrate that human ESCs, iPSCs and ASCs can be maintained in the same defined xeno-free medium formulation for a prolonged period of time while maintaining their characteristics, demonstrating the applicability of the simplified xeno-free medium formulation for the production of clinical-grade stem cells. The basic xeno-free formulation described herein has the potential to be further optimized for specific applications relating to establishment, expansion and differentiation of various stem cell types.
    Document Type:
    Reference
    Product Catalog Number:
    AB5032
    Product Catalog Name:
    Anti-Neural Cell Adhesion Molecule Antibody
  • «
  • <
  • 1
  • >
  • »