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N2 media

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N2 media is a specialized cell culture supplement designed to support the growth and maintenance of neural cells. It provides a defined, serum-free environment that promotes the survival and differentiation of various types of neural cells, including neurons, astrocytes, and oligodendrocytes. The core function of N2 media is to create an optimized culture condition for the study and manipulation of neural cell biology.

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Lab products found in correlation

Knockout serum replacement (ksr), by Thermo Fisher Scientific (2 mentions) Fibroblast growth factor 2 (fgf2), by Thermo Fisher Scientific (2 mentions) Mem non essential amino acid, by Thermo Fisher Scientific (2 mentions) β mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Ldn193189, by Thermo Fisher Scientific (2 mentions) Sb431542, by Abcam (2 mentions) Knockout dmem, by Thermo Fisher Scientific (2 mentions) Y 27632, by Bio-Techne (2 mentions) L glutamine, by Thermo Fisher Scientific (2 mentions) Neurobasal media, by Thermo Fisher Scientific (1 mentions) 257 nt cf, by R&D Systems (1 mentions) 212 gd cf, by R&D Systems (1 mentions) Sb431542, by Bio-Techne (1 mentions)

3 protocols using n2 media

1

Neural Induction and Differentiation

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Neural induction was performed as previously reported33 . Briefly, cells were dissociated to single cells using Accutase and plated on gelatin for 10 minutes to remove MEFs. Non-adherent cells were collected and plated on Geltrex-treated dishes at a density of 150-200k cells per well of a 24-well plate in the presence of MEF-conditioned hESC media containing 10 ng/ml of FGF-2 (Life Tech) and 10 uM of Y-27632 (Tocris). Neural differentiation was initiated when cells were confluent using KSR media containing 820 ml of Knockout DMEM (Life Tech), 150 ml Knockout Serum Replacement (Life Tech), 1 mM L-glutamine (Life Tech), 100 uM MEM non-essential amino acids (Life Tech), and 0.1 mM beta- mercaptoethanol (Life Tech) to inhibit SMAD signaling, 100 nM of LDN-193189 (Cat. no. ab142186, Abcam) and 5 uM of SB431542 (Cat. No. 13031, Cayman Chemical) were added on Days 0 through 9. Cells were fed daily, and N2 media (Life Tech) was added in increasing 25% increments every other day starting on Day 4 (100% N2 on Day 10).
Choi J., Lee S., Clement K., Mallard W., Tagliazucchi G.M., Lim H., Choi I.Y., Ferrari F., Tsankov A., Pop R., Lee G., Rinn J., Meissner A., Park P.J, & Hochedlinger K. (2015). A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs. Nature biotechnology, 33(11), 1173-1181.
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2

Differentiation of iPSCs into Neurons

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iPSCs stably expressing TetO-Ngn2-Neo and reverse tetracycline-controlled transactivator (rtTA) were plated at a density of 40,000 cells cm−2 with rock inhibitor Y27632 (Stemgent, 04-0012). Day 1 cells were differentiated in N2 media (Life Technologies) supplemented with 10 μM SB431542 (Tocris, 1614), 2 μM XAV939 (Stemgent, 04-00046) and 100 nM LDN-193189 (Stemgent, 04-0074) along with doxycycline hyclate (2 μg mL−1). Day 2 media was N2+SB/XAV/LDN/doxycycline hyclate and differentiation media were as previously described. On day 3, cell differentiation was continued in neurobasal media (Life Technologies) supplemented with B27 (50X, Thermo Scientific), brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CTNF), glial cell-derived neurotrophic factor (GDNF) (R&D Systems 248-BD/CF, 257-NT/CF, and 212-GD/CF at 10 ng mL−1) and doxycyclinehyclate (2 μg mL−1).
Pintacuda G., Hsu Y.H., Tsafou K., Li K.W., Martín J.M., Riseman J., Biagini J.C., Ching J.K., Mena D., Gonzalez-Lozano M.A., Egri S.B., Jaffe J., Smit A.B., Fornelos N., Eggan K.C, & Lage K. (2023). Protein interaction studies in human induced neurons indicate convergent biology underlying autism spectrum disorders. Cell Genomics, 3(3), 100250.
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3

Neural Induction and Differentiation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Neural induction was performed as previously reported33 . Briefly, cells were dissociated to single cells using Accutase and plated on gelatin for 10 minutes to remove MEFs. Non-adherent cells were collected and plated on Geltrex-treated dishes at a density of 150-200k cells per well of a 24-well plate in the presence of MEF-conditioned hESC media containing 10 ng/ml of FGF-2 (Life Tech) and 10 uM of Y-27632 (Tocris). Neural differentiation was initiated when cells were confluent using KSR media containing 820 ml of Knockout DMEM (Life Tech), 150 ml Knockout Serum Replacement (Life Tech), 1 mM L-glutamine (Life Tech), 100 uM MEM non-essential amino acids (Life Tech), and 0.1 mM beta- mercaptoethanol (Life Tech) to inhibit SMAD signaling, 100 nM of LDN-193189 (Cat. no. ab142186, Abcam) and 5 uM of SB431542 (Cat. No. 13031, Cayman Chemical) were added on Days 0 through 9. Cells were fed daily, and N2 media (Life Tech) was added in increasing 25% increments every other day starting on Day 4 (100% N2 on Day 10).
Choi J., Lee S., Clement K., Mallard W., Tagliazucchi G.M., Lim H., Choi I.Y., Ferrari F., Tsankov A., Pop R., Lee G., Rinn J., Meissner A., Park P.J, & Hochedlinger K. (2015). A comparison of genetically matched cell lines reveals the equivalence of human iPSCs and ESCs. Nature biotechnology, 33(11), 1173-1181.
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