Essential 8 flex

Manufactured by Thermo Fisher Scientific

The Essential 8 Flex is a cell culture media designed for the maintenance and expansion of human pluripotent stem cells. It provides a defined, animal component-free, and xeno-free environment for cell culture.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Essential 8 (61 citations) Essential 8 Flex medium (52 citations) Essential 8 Flex Medium Kit (6 citations) Essential 8 Flex media (5 citations) Essential 8 Flex medium (E8; (4 citations) Essential 8TM Flex medium (2 citations) Essential 8 Flex Medium (E8f) (2 citations) Essential 8 Flex Media Kit (2 citations)

6 protocols using essential 8 flex

Hepatocyte Differentiation of Stem Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human embryonic stem cell (hESC) line H9 (WA09) was purchased from WiCell Research Institute (Madison, 15 WI) and kept according to supplier’s instructions as feeder-free cultures on Matrigel (BD biosciences)-coated plates in Essential 8 (or Essential 8 Flex) (Thermo fisher). BJ1 iPSCs were made in-house and kept on Matrigel-coated plates at ±8.75 × 10⁴ cells/cm² in mTeSR medium (Stem Cell Technologies)^{65 (link)}. Cells were differentiated towards the hepatocyte lineage as described by our group before^{65 (link)}, with some optimizations. Briefly, single-cell suspensions of H9/ BJ1 cells (using accutase) were plated on Matrigel-coated plates at ±8.75 × 10⁴ cells/cm² in mTeSR medium (Stem Cell Technologies). When cells reached 70–80% confluency, differentiation was started using the previously described cytokine regimes and was stopped after 20 days of differentiation. All cytokines were purchased from Peprotech (NJ). Differentiation medium was supplemented with 0.6% DMSO during the first 12 days of the culture and with 2% DMSO during the last 8 days of differentiation. Use of embryonic stem cells and iPSCs for research was approved by the “Commissie Medische Ethiek,” UZ KU Leuven/Onderzoek U.Z. Gasthuisberg, Herestraat 49, B 3000 Leuven, under file number S52426

Boon R., Kumar M., Tricot T., Elia I., Ordovas L., Jacobs F., One J., De Smedt J., Eelen G., Bird M., Roelandt P., Doglioni G., Vriens K., Rossi M., Vazquez M.A., Vanwelden T., Chesnais F., El Taghdouini A., Najimi M., Sokal E., Cassiman D., Snoeys J., Monshouwer M., Hu W.S., Lange C., Carmeliet P., Fendt S.M, & Verfaillie C.M. (2020). Amino acid levels determine metabolism and CYP450 function of hepatocytes and hepatoma cell lines. Nature Communications, 11, 1393.

+ Open protocol

+ Expand

Rhesus CD47 Overexpressing iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human B2M⁻^/⁻CIITA⁻^/⁻ iPSCs overexpressing rhesus CD47 (rhCD47) were cultured on diluted feeder-free Matrigel (hESC qualified, BD Biosciences)–coated 6-well plates in Essential 8 Flex medium (Thermo Fisher) and transduced to express Fluc as described above.

Deuse T., Hu X., Agbor-Enoh S., Jang M.K., Alawi M., Saygi C., Gravina A., Tediashvili G., Nguyen V.Q., Liu Y., Valantine H., Lanier L.L, & Schrepfer S. (2021). The SIRPα–CD47 immune checkpoint in NK cells. The Journal of Experimental Medicine, 218(3), e20200839.

+ Open protocol

+ Expand

Cardiomyocyte Differentiation of RBM20 Mutant iPSCs

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Parental iPSCs and iPSCs harboring the homozygous P633L or R634Q mutation in RBM20 were previously generated and characterized^{24 (link)}. Cells were maintained on vitronectin (A31804, ThermoFisher) coated plates with Essential 8™ Flex (A2858501, ThermoFisher) medium and passaged with Versene (15040066, ThermoFisher). Cardiomyocyte differentiation was initiated by addition of 8 μM CHIR99021 (72054, STEMCELL Technologies) in RPMI-1640 medium supplemented with B27 without Insulin (RPMI-Insulin, A1895601, ThermoFisher). After 24 h, 1 volume of RPMI-Insulin was added and after 72 h, medium was changed to RPMI-Insulin with 2 μM Wnt-C59 (5148, Tocris). At day 5 and 7, medium was changed to RPMI-Insulin and at day 9 to RPMI with full B27 supplement (RPMI+Insulin, 17504044, ThermoFisher). At day 11, medium was changed to RPMI+Insulin without glucose and addition of 5 mM DL-lactate. At day 14, RPMI+Insulin was added and at day 16, cells were passaged with TrypLE10x (A1217701, ThermoFisher) and RPMI+Insulin supplemented with 10% knock-out serum replacement (10828028, ThermoFisher) and 1.66 μM Thiazovivin (72252, StemCell Technologies). One day after passaging, the medium was changed to RPMI+Insulin with subsequent medium exchange every 3 days. Passaging was done every 2–3 weeks.

Grosch M., Schraft L., Chan A., Küchenhoff L., Rapti K., Ferreira A.M., Kornienko J., Li S., Radke M.H., Krämer C., Clauder-Münster S., Perlas E., Backs J., Gotthardt M., Dieterich C., van den Hoogenhof M.M., Grimm D, & Steinmetz L.M. (2023). Striated muscle-specific base editing enables correction of mutations causing dilated cardiomyopathy. Nature Communications, 14, 3714.

+ Open protocol

+ Expand

Culturing and Maintaining Human iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human iPSCs (hiPSC-L1) were cultured on six well plates coated with growth factor-reduced Matrigel (Corning) in an incubator at 37°C, 5% CO₂. Stem cells were maintained in Essential 8 Flex medium (E8 Flex; Thermo Fisher Scientific) containing 1% penicillin/streptomycin until ∼70% confluency was reached, at which point cells were split into new wells using ReLeSR passaging reagent (Stem Cell Technologies) with 2 µM ROCK inhibitor (Thiazovivin/TZV; Selleck Chemicals) added to prevent cells from undergoing apoptosis while in suspension. All hiPSC lines were periodically validated for pluripotency and genomic stability.

Wasserman A.H., Huang A.R., Lewis-Israeli Y.R., Dooley M.D., Mitchell A.L., Venkatesan M, & Aguirre A. (2022). Oxytocin promotes epicardial cell activation and heart regeneration after cardiac injury. Frontiers in Cell and Developmental Biology, 10, 985298.

+ Open protocol

+ Expand

Culturing Human Embryonic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human ESCs (WA25 hESCs and genomically edited cell lines based on the WA25 background, passage 13–55) were cultured in Essential 8 Flex (E8f) medium (Thermo Fisher) supplemented with 100 µg/mL Normocin (Invivogen) (hereafter, E8fn medium) on truncated recombinant human Vitronectin-N (Thermo Fisher)-coated Nunclon Delta surface-treated 6-well plates (Thermo Fisher) according to an established protocol^{58 (link)}. At 60%–80% confluency or every 3 or 4 days, the cells were passaged at a split ratio of 1:100–1:10 into 2–4 wells of a 6-well plate using 0.5 mM Ethylenediaminetetraacetic acid (EDTA) in Dulbecco’s phosphate-buffered saline (DPBS). 1X RevitaCell (Thermo Fisher) was supplemented in the E8fn medium for 1 d after passaging for increased viability. The WA25 hES cell line was acquired from the WiCell Research Institute. For additional validation and testing information, refer to the cell line webpage: https://www.wicell.org/home/stem-cell-lines/catalog-of-stem-cell-lines/wa25.cmsx.

Nie J., Ueda Y., Solivais A.J, & Hashino E. (2022). CHD7 regulates otic lineage specification and hair cell differentiation in human inner ear organoids. Nature Communications, 13, 7053.

+ Open protocol

+ Expand

Inducing Hepatocyte-like Cells from iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

The normal donor SIGi001 iPSC0028 was purchased from Sigma. Both cell lines were genetically modified, as previously described, 9, (link)18 (link) to incorporate a doxycycline inducible cassette encoding for HNF1A, PROX1, and FOXA3 with or without PGC1A, SIRT1, and activated AMPK (respectively named HC3x and HC6x). PSCs were maintained according to supplier's instructions as feeder-free cultures on Matrigel (BD biosciences)-coated plates in Essential 8 Flex (Thermo Fisher). Cells were differentiated toward the hepatocyte lineage as described by our group before. 9 (link)

Moreno-Torres M., Kumar M., García-Llorens G., Quintás G., Tricot T., Boon R., Tolosa L., Toprakhisar B., Chesnais F., Verfaillie C, & Castell J.V. (2022). A Novel UPLC-MS Metabolomic Analysis-Based Strategy to Monitor the Course and Extent of iPSC Differentiation to Hepatocytes. Journal of proteome research, 21(3).

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!