Enstem a neural expansion medium

Manufactured by Merck Group

ENStem-A is a neural expansion medium developed by Merck Group. It is designed to support the growth and maintenance of neural stem cells in vitro.

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Lab products found in correlation

Fibroblast growth factor 2 (fgf2), by Merck Group (1 mentions) Pen strep, by Thermo Fisher Scientific (1 mentions) Acutase, by Merck Group (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Matrigel, by BD (1 mentions)

2 protocols using enstem a neural expansion medium

Generation of Self-Renewing Neural Progenitor Cells

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The RG and TCL cells were plated at a density of 300 cells per ml on 24 well plates in ENStem-A neural expansion medium with FGF2 at 20 ng/ml (Millipore), L-glutamine 2 mM and PenStrep 1× (Gibco) and grown for 14 days at 37°C, 5% CO2 in a humidified atmosphere. The neurospheres were collected and plated in the same media on laminin-coated tissue culture plates for 24-48 hours (in order to convert the neurospheres into the cell monolayer) at 37°C, 5% CO2 in a humidified atmosphere. Acutase (Millipore) cell detachment was applied and the neurosphere formation process repeated again to produce self-renewal cell culture. The self-renewing cells were named LC26-R-2^nd nsphr and LC26-RTL(170)-2^nd nsphr, LCAS-R-2^nd nsphr and LCAS-RTL(138)-2^nd nsphr, respectively.

Malchenko S., Sredni S.T., Boyineni J., Bi Y., Margaryan N.V., Guda M.R., Kostenko Y., Tomita T., Davuluri R.V., Velpula K., Hendrix M.J, & Soares M.B. (2018). Characterization of brain tumor initiating cells isolated from an animal model of CNS primitive neuroectodermal tumors. Oncotarget, 9(17), 13733-13747.

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Differentiation of Human Neural Progenitor Cells

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Commercially available human neural progenitor cells (hNPCs) derived from NIH approved H9 stem cell line with normal female karyotype were obtained (ArunA Biomedical, Athens, GA, now available as ENStem-A hNPCs from Millipore, Billerica, MA) and expanded 8–10 passages in ENStem-A™ neural expansion medium (Millipore, Billerica, MA). For long-term differentiation experiments, hNPC culture protocols were modified from protocols developed by ArunA Biomedical (Athens, GA). At passages 8–10, cells were plated at a density of 200,000 cells/ml in 35mm dishes coated with Matrigel (BD Biosciences, Franklin Lakes, NJ) diluted 1:200 in DMEM/F-12. Cells were allowed to attach and proliferate for 24 hours; then the medium was replaced with Hyclone neuronal differentiation medium (Thermo Fisher Scientific, Waltham, MA) in the absence of growth factor to initiate differentiation. Cells were maintained in differentiation conditions for up to 21 days, with half of the medium refreshed every two to three days and whole medium refreshed weekly.

Wegner S.H., Park J.J., Workman T., Hermsen S.A., Wallace J., Stanaway I.B., Kim H.Y., Griffith W.C., Hong S, & Faustman E.M. (2019). Anchoring a dynamic in vitro model of human neuronal differentiation to key processes of early brain development in vivo. Reproductive toxicology (Elmsford, N.Y.), 91, 116-130.

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