Gem21

Manufactured by Gemini Bio

Sourced in United States

The Gem21 is a versatile laboratory instrument designed for cell culture and molecular biology applications. It features precise temperature control, uniform heating, and advanced programming capabilities to support a range of experimental protocols.

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Lab products found in correlation

Close spelling variants of this product

Gem21 NeuroPlex (14 citations) Gem21 NeuroPlex Serum-Free Supplement (4 citations) B27 Gem21 Neuroplex (2 citations) Gem21 supplement (2 citations)

4 protocols using gem21

Modulating Sox2 Expression in Cancer Cells

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Skov-3, Pa-1, OVCAR3, OVCAR4, and OVCAR5 cell lines were obtained from ATCC. A2780 parental cells (IP2) and cisplatin resistant cells (CP20) were generously donated by Dr. Charles Landen (University of Virginia). Cells were grown in RPMI (Skov-3, A2780, OVCAR4) or DMEM (Pa-1, OVCAR5) media containing 10% fetal bovine serum (FBS, Atlanta Biologicals) and antibiotic/antimycotic supplements (Invitrogen). OVCAR3 cells were grown in RPMI with 20% FBS and 0.01 mg/mL of bovine insulin (Sigma). Normal human astrocytes (NHA, Lonza) were cultured in AGM media, and immortalized neural progenitor cells (NPC, Millipore) were propagated in DMEM/F12 supplemented with EGF, FGF and Gem21 (Gemini Bio-Products). Stable polyclonal cell lines with either forced expression of Sox2 (GeneCopoeia), or shRNA against Sox2 (Sigma), were created by lentiviral transduction followed by puromycin selection. Cells with inducible Sox2 expression were generated using lentivirus harboring a tetracycline-inducible Sox2 construct (GeneCopoeia) followed by selection with blasticidin. Sox2 expression was induced in this latter cell line with 1 μg/ml doxycycline. In a pilot experiment, dox-induced Sox2 expression was measured at multiple time points, and based on these data, all further dox treatments were conducted at 96 h. Modulation of Sox2 expression in these various cell models was confirmed by immunoblotting.

Dorsett K.A., Jones R.B., Ankenbauer K.E., Hjelmeland A.B, & Bellis S.L. (2019). Sox2 promotes expression of the ST6Gal-I glycosyltransferase in ovarian cancer cells. Journal of Ovarian Research, 12, 93.

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Transplantation of Human Neural Progenitor Cells

Cited 2 times

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Human NPCs were obtained according to NIH Ethical Guidelines and have been fully described in previous studies^{21 (link)–23 (link, link)}. Briefly, NPCs were grown for 13 doublings in ultra-low attachment culture flasks under feeder-free conditions in serum and xeno-free Eagle’s essential medium (Hyclone, Logan, UT, USA), supplemented with Gem21 (Gemini Bio-Products, Sacramento, CA, USA), epidermal growth factor (Peprotech, Rocky Hill, NJ, USA), basic fibroblast growth factor (Peprotech), transforming growth factor alpha (Peprotech), insulin-like growth factor I (Peprotech), leukemia inhibitory factor (Millipore, Temecula, CA, USA), calcium chloride (Fisher Scientific, Waltham MA, USA), Glutamax (Invitrogen, Carlsbad, CA, USA), non-essential amino acids (Hyclone), and an N2 supplement (Invitrogen), all of which were added at proprietary concentrations. The dead NPCs were implemented as a negative control for any paracrine effects from the live NPC inoculations. The dead NPCs were obtained from a live NPC population and then were placed without cryoprotectant into a −20°C freezer for 30 min to freeze kill the cells, and then stored in −80°C freezer until use. The suspended dead NPCs in culture medium were thawed for 1 h and then used immediately for transplantation in sHW mutants.

Tierney W.M., Uhlendorf T.L., Lemus A.J., Ortega B.A., Magaña J., Ochoa J., Van Trigt W., Cruz A., Kopyov A., Kopyov O.V, & Cohen R.W. (2020). Transplanted Human Neural Progenitor Cells Attenuate Motor Dysfunction and Lengthen Longevity in a Rat Model of Ataxia. Cell Transplantation, 29, 0963689720920275.

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Microglia-like Cells from iPSCs

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We derived microglia-like cells from iPSCs as previously described (Muffat et al., 2016 (link)). Briefly, iPSCs on MEFs-coated plates were dissociated with Collagenase IV (Thermo Fisher Scientific). iPSCs were then resuspended in MGD media [Neurobasal media supplemented with 0.5X Gem21 (Gemini Bio-products), 0.5X Neuroplex N2 (Gemini Bio-products), 0.2% Albumax I (Thermo Fisher Scientific), 5 mM sodium chloride (Sigma-Aldrich), 1X sodium pyruvate, 1X P/S, 1X GlutaMAX, 3.5 ng/ml biotin (Sigma-Aldrich), 10 μM ascorbic acid (Sigma-Aldrich) and 1.7% lactic syrup (Sigma-Aldrich)] with 10 ng/ml IL-34 (Peprotech) and 10 ng/ml M-CSF1(Peprotech)], and cultured in ultra-low attachment 6-well plates (Corning). Once the phase-bright neutralized spheroids and cystic bodies appeared, Embryoid bodies were gently triturated to shear off cells of interest, and supernatants were transferred to a single well of Primaria 6-well plate (Corning). Attached cells showed morphological characteristics of microglia and microglia precursors. Cells from 6 consecutive productions were pooled and purified by FACS using CD11b antibody (Miltenyi Biotec). Collected cells were further maintained in MGD media with 100 ng/ml IL-34 and 5 ng/ml M-CSF for all experiments.

Lin Y.T., Seo J., Gao F., Feldman H.M., Wen H.L., Penney J., Cam H.P., Gjoneska E., Raja W.K., Cheng J., Rueda R., Kritskiy O., Abdurrob F., Peng Z., Milo B., Yu C.J., Elmsaouri S., Dey D., Ko T., Yankner B.A, & Tsai L.H. (2018). APOE4 causes widespread molecular and cellular alterations associated with Alzheimer’s disease phenotypes in human iPSC-derived brain cell types. Neuron, 98(6), 1141-1154.e7.

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Induced Pluripotent Stem Cell Differentiation

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The iPSC medium was changed to N2 medium: DMEM/F12 with 1× N2 (Invitrogen) and dual SMAD inhibitors: 1 μM dorsomorphin (Tocris) and 10 μM SB431542 (Stemgent). Colonies in suspension as Embryoid Bodies (EBs) after 5 day at 90 rpm in N2 medium were plated on matrigel-coated plates with NGF medium: DMEM/F12 with 0.5× N2, 0.5× Gem21 (Gemini Bioproducts), 20 ηg/mL FGF2 and 1% Pen/Strep. Rosettes containing neural progenitor cells (NPCs) were dissociated and plated in poly-ornithine (10 μg/mL, Sigma) and Laminin (2.5 μg/mL, ThermoFisher) coated plate. NPCs expansion used NGF medium.

Correa Leite P.E., de Araujo Portes J., Pereira M.R., Russo F.B., Martins-Duarte E.S., Almeida dos Santos N., Attias M., Barrantes F.J., Baleeiro Beltrão-Braga P.C, & de Souza W. (2020). Morphological and biochemical repercussions of Toxoplasma gondii infection in a 3D human brain neurospheres model. Brain, Behavior, & Immunity - Health, 11, 100190.

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