Log In Sign up
The largest database of trusted experimental protocols

Stemflex medium

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States, Japan

StemFlex medium is a defined, serum-free culture medium designed for the maintenance and expansion of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) in a feeder-free culture system.

Automatically generated - may contain errors

Lab products found in correlation

Matrigel, by Corning (41 mentions) Geltrex, by Thermo Fisher Scientific (21 mentions) Relesr, by STEMCELL (20 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (18 mentions) Dmem f12, by Thermo Fisher Scientific (11 mentions) Gentle cell dissociation reagent (gcdr), by STEMCELL (9 mentions) Versene, by Thermo Fisher Scientific (8 mentions) Glutamax, by Thermo Fisher Scientific (8 mentions) Cytotune ips 2.0 sendai reprogramming kit, by Thermo Fisher Scientific (7 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (7 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions) Revitacell, by Thermo Fisher Scientific (6 mentions) Geltrex matrix, by Thermo Fisher Scientific (6 mentions) Cytotune ips 2.0 sendai reprogramming, by Thermo Fisher Scientific (6 mentions) Tryple, by Thermo Fisher Scientific (5 mentions) Retinoic acid, by Merck Group (5 mentions) Matrigel, by BD (5 mentions) Vitronectin, by Thermo Fisher Scientific (5 mentions) Versene solution, by Thermo Fisher Scientific (5 mentions) Pen strep, by Thermo Fisher Scientific (5 mentions)

Spelling variants of this product

StemFlex (41 citations) StemFlex Basal medium (4 citations) StemFlex medium kit (4 citations) Gibco™ StemFlex™ Medium (2 citations)

174 protocols using stemflex medium

1

Generation and Characterization of iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
iPSCs were generated from fibroblasts at the Harvard Stem Cell Institute iPS core, using the CytoTune-iPS 2.0 Sendai Reprogramming Kit (ThermoFisher #A16517). Eight days after transduction, iPSCs were replated and maintained in StemFlex medium (ThermoFisher #A3349401) on Cultrex (R&D #3434-001-02). Emerging iPSC colonies were manually picked and plated between 7/3/18–7/9/18 and expanded in StemFlex medium on Cultrex at 37°C in 5% CO2. Cell were tested for mycoplasma and cryopreserved in 5 cryovials. One vial was thawed to test for recovery, and cells were used for pluipotency immunocytochemistry, pluripotency RT-qPCR, embroid body RT-qPCR for trilineage differentiation, and sent to WiCell for G-banded karyotyping. The iPSCs were subsequently cultured in StemFlex medium on geltrex (ThermoFisher #A1413302) and passaged every 5–7 days using Gentle Cell Dissociation Reagent (StemCell #07174) or ReLeSR (StemCell #05872).
Chen P.F., Chen T., Forman T.E., Swanson A.C., O’Kelly B., Dwyer S.A., Buttermore E.D., Kleiman R., Carrington S.J., Lavery D.J., Swanson L.C., Olson H.E, & Sahin M. (2021). Generation and characterization of human induced pluripotent stem cells (iPSCs) from three male and three female patients with CDKL5 Deficiency Disorder (CDD). Stem cell research, 53, 102276.
+ Open protocol
+ Expand
2

Cardiomyocyte Differentiation of hiPSCs

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Subclones from two control hiPSC lines (LUMC0020iCTRL (Zhang et al., 2014 (link)) [LUMC20]; LUMC0099iCTRL [LUMC99]), and two hiPSC lines derived from patients diagnosed with HCM (LUMC0033iMyBPC [HCM1]; LUMC0035iMyBPC [HCM3] (Birket et al., 2015 (link))) were maintained either in Essential 8 or StemFlex Medium (both Gibco). One day prior to differentiation (d-1), the hiPSCs were harvested using TrypLE Select (Gibco) and plated onto Matrigel-coated wells of a 12-well cell culture plate, either in Essential 8 Medium containing RevitaCell Supplement (1:200 dilution; Gibco) or StemFlex Medium at 3.9 × 104/cm2. The hiPSCs were differentiated into cardiomyocytes either using the Pluricyte Cardiomyocyte Differentiation Kit (NCardia) according to the manufacturer's instructions, or in a modified BPEL medium (Elliott et al., 2011 (link)) supplemented with small molecules. Specifically, 5 μM CHIR99021 (Axon Medchem) from day 0 to day 2 of differentiation and 5 μM XAV939 + 0.25 μM IWP-L6 (AbMole) from differentiation day 2 to day 4. The hiPSC-CMs were maintained in Medium C (NCardia) until differentiation day 20–21 (LUMC20 and LUMC99), or the modified BPEL medium until differentiation day 14 or 17 (HCM1 and HCM3), and then dissociated as previously described (van den Berg et al., 2014 ).
van den Brink L., Brandão K.O., Yiangou L., Mol M.P., Grandela C., Mummery C.L., Verkerk A.O, & Davis R.P. (2020). Cryopreservation of human pluripotent stem cell-derived cardiomyocytes is not detrimental to their molecular and functional properties. Stem cell research, 43, 101698.
+ Open protocol
+ Expand
3

Cardiomyocyte Differentiation of hiPSCs

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Subclones from two control hiPSC lines (LUMC0020iCTRL (Zhang et al., 2014 (link)) [LUMC20]; LUMC0099iCTRL [LUMC99]), and two hiPSC lines derived from patients diagnosed with HCM (LUMC0033iMyBPC [HCM1]; LUMC0035iMyBPC [HCM3] (Birket et al., 2015 (link))) were maintained either in Essential 8 or StemFlex Medium (both Gibco). One day prior to differentiation (d-1), the hiPSCs were harvested using TrypLE Select (Gibco) and plated onto Matrigel-coated wells of a 12-well cell culture plate, either in Essential 8 Medium containing RevitaCell Supplement (1:200 dilution; Gibco) or StemFlex Medium at 3.9 × 104/cm2. The hiPSCs were differentiated into cardiomyocytes either using the Pluricyte Cardiomyocyte Differentiation Kit (NCardia) according to the manufacturer's instructions, or in a modified BPEL medium (Elliott et al., 2011 (link)) supplemented with small molecules. Specifically, 5 μM CHIR99021 (Axon Medchem) from day 0 to day 2 of differentiation and 5 μM XAV939 + 0.25 μM IWP-L6 (AbMole) from differentiation day 2 to day 4. The hiPSC-CMs were maintained in Medium C (NCardia) until differentiation day 20–21 (LUMC20 and LUMC99), or the modified BPEL medium until differentiation day 14 or 17 (HCM1 and HCM3), and then dissociated as previously described (van den Berg et al., 2014 ).
van den Brink L., Brandão K.O., Yiangou L., Mol M.P., Grandela C., Mummery C.L., Verkerk A.O, & Davis R.P. (2020). Cryopreservation of human pluripotent stem cell-derived cardiomyocytes is not detrimental to their molecular and functional properties. Stem cell research, 43, 101698.
+ Open protocol
+ Expand
4

Fibroblast-derived iPSC Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols
Patient skin fibroblasts were obtained from Coriell Cell Repositories (GM11097) and cultured in DMEM (Thermo Fisher Scientific), supplemented with 10% fetal bovine serum (FBS), 100 units/ml penicillin, and 100 μg/ml streptomycin in a humidified incubator with 5% CO2 at 37 °C. TRNDi004-I iPSCs were cultured in StemFlex medium (Thermo Fisher Scientific) on Geltrex (Thermo Fisher Scientific)-coated plates. The cells were maintained at 5% CO2, 5% O2 at 37 °C and the cells were passaged with 0.5 mM ethylenediaminetetraacetic acid (EDTA) when colonies were approximately 70% confluent.
Baskfield A., Li R., Beers J., Zou J., Liu C, & Zheng W. (2019). Generation of an induced pluripotent stem cell line (TRNDi004-I) from a Niemann-Pick disease type B patient carrying a heterozygous mutation of p.L43_A44delLA in the SMPD1 gene. Stem cell research, 37, 101436.
+ Open protocol
+ Expand
5

Culturing and Maintaining Human iPSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
iPSCs are cultured in StemFlex medium (ThermoFisher) on Matrigel (Millipore). Pluripotent cultures are maintained by manually picking colonies with overt pluripotent morphology (small tightly packed cells, smooth colony borders, low cytoplasmic to nuclear ratio) using a sterile pulled glass pipet under an inverted light microscope into sterile PBS, centrifuged to remove the PBS, and transferred to a new Matrigel-coated plate in StemFlex medium. Cultures are expanded before experiments by passaging using ReLeSR (STEMCELL Technologies) as per the manufacturer’s instructions into Matrigel coated plates with StemFlex medium.
Singh T., Jiao Y., Ferrando L.M., Yablonska S., Li F., Horoszko E.C., Lacomis D., Friedlander R.M, & Carlisle D.L. (2021). Neuronal mitochondrial dysfunction in sporadic amyotrophic lateral sclerosis is developmentally regulated. Scientific Reports, 11, 18916.
+ Open protocol
+ Expand
6

Generation and Maintenance of Human iPSCs and Mouse Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
Two human iPSC lines, BJFF.6 from a male donor and AN1.1 from a female donor, were generated at the Genome Engineering & Stem Cell Center (GESC) at Washington University, St Louis, MO, USA. iPSCs were maintained on Matrigel (Corning)-coated plates in Stem-Flex medium (Thermo Fisher Scientific). In addition, four mouse cell lines – NIH3T3, MC3T3-E1, CT26 and 4T1 – were obtained from the ATCC and cultured in the ATCC-recommended media. All cell lines were grown in a humidified incubator set at 5% CO2 and 37°C.
Chen Y.H., Connelly J.P., Florian C., Cui X, & Pruett-Miller S.M. (2023). Short tandem repeat profiling via next-generation sequencing for cell line authentication. Disease Models & Mechanisms, 16(10), dmm050150.
+ Open protocol
+ Expand
7

Generation of iPS-derived Neurons

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The generation of cell lines from human iPS cells was approved by the institutional review board (IRB) of Brigham & Women’s Hospital (IRB protocol 2015P001676). iPSCs were generated from peripheral blood mononuclear cells (PBMCs) from the Religious Order Study (ROS) and Memory and Aging Project (MAP) cohort using the Sendai virus reprogramming method as previously described25 (link). iPS cells underwent a rigorous quality check procedure that includes a sterility check, mycoplasma testing, karyotyping and pluripotency assays performed by the New York Stem Cell Foundation (NYSCF). iPS cells were maintained using StemFlex Medium (Thermo Fisher Scientific). For this study, two cell lines (one male and one female) were used for induced neuron differentiation.
Clark L.E., Clark S.A., Lin C., Liu J., Coscia A., Nabel K.G., Yang P., Neel D.V., Lee H., Brusic V., Stryapunina I., Plante K.S., Ahmed A.A., Catteruccia F., Young-Pearse T.L., Chiu I.M., Llopis P.M., Weaver S.C, & Abraham J. (2021). VLDLR and ApoER2 are receptors for multiple alphaviruses. Nature, 602(7897), 475-480.
+ Open protocol
+ Expand
8

Generation of Induced Neurons from iPSCs

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The generation of cell lines from human induced pluripotent stem cells (iPSCs) was approved by the institutional review board (IRB) of Brigham & Women’s Hospital (IRB protocol #2015P001676). iPSCs were generated from peripheral blood mononuclear cells (PBMCs) from the Religious Order Study (ROS) and Memory and Aging Project (MAP) cohort using the Sendai virus reprogramming method as previously described25 (link). iPSCs underwent a rigorous quality check procedure that includes a sterility check, mycoplasma testing, karyotyping, and pluripotency assays performed by the New York Stem Cell Foundation (NYSCF). iPSCs were maintained using StemFlex™ Medium (Thermo Fisher Scientific). For this study, two cell lines (one male and one female) were used for induced neuron differentiation.
Clark L.E., Clark S.A., Lin C., Liu J., Coscia A., Nabel K.G., Yang P., Neel D.V., Lee H., Brusic V., Stryapunina I., Plante K.S., Ahmed A.A., Catteruccia F., Young-Pearse T.L., Chiu I.M., Llopis P.M., Weaver S.C, & Abraham J. (2021). VLDLR and ApoER2 are receptors for multiple alphaviruses. Nature, 602(7897), 475-480.
+ Open protocol
+ Expand
9

Cerebral and Ventral Organoid Generation

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
Male (XY) and female (XX) stem cells (male - H1 (WiCell WA01) and female - H9 (WiCell WA09)) were approved for use in the project by the UK Stem Cell Bank Steering Committee. Cells were cultured on Matrigel® (Corning, 356234) – coated plates and maintained in StemFlex™ medium (Thermo Fisher, A3349401). We used the commercial kit (STEMdiff™ Cerebral Organoid Kit, StemCell Technologies 08570) with previously described modifications to increase embryoid body surface area52 (link)
, namely microscaffolds (18000 cells seeded) or smaller EBs (2000 cells seeded). Organoids were transferred to the orbital shaker at 57 rpm (f25 cm) at 15d (±1 day, depending on the morphology). Matrigel® was removed manually between days 14-17, after visual inspection. Organoids were transferred to IDM+A supplemented with diluted Matrigel® on day 30 and kept at those conditions until fixation. Ventral organoids were generated as previously described53 (link)
, with ventralizing factors (2.5 μM IWP-2, Sigma, I0536; 100 nM SAG, Sigma, 566661) supplemented in the neural induction (NI) medium for 2 days.
Kelava I., Chiaradia I., Pellegrini L., Kalinka A.T, & Lancaster M.A. (2022). Androgens increase excitatory neurogenic potential in human brain organoids. Nature, 602(7895), 112-116.
+ Open protocol
+ Expand
10

Differentiation of iPSCs to Cardiomyocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell reprogramming into iPSC lines was previously described by Rovina et al. [15 (link)]. Briefly, iPSCs were seeded in Matrigel (Corning, 356,230) coated 6 well plates in StemFlex medium (ThermoFisher, A3349401) supplemented with RevitaCell (ThermoFisher, A2644501). On Day 0, medium was replaced with BPEL medium and cells were treated with 6 mM CHIR99021 (Bertin bioreagent, 13,122), 100 µg/ml Activin A (MiltenyiBiotec, 130–115-008), and 100 µg/ml BMP4 (R&D Systems, 5020-BP). On Day 3, cells were treated with 10 mM XAV939 (R&D Systems,3748), 100 µg/ml BMP4 and 2 mM Retinoic acid (Sigma Aldrich, R2625). Medium is changed on Day6 with BPEL + 30 ng/ml BMP4 + 1 µM Retinoic acid. After 3 days, cells were replated on fibronectin coated plates with BPEL medium supplemented with 20 mM SB 431542 (R&D Systems,161).
Soussi S., Savchenko L., Rovina D., Iacovoni J.S., Gottinger A., Vialettes M., Pioner J.M., Farini A., Mallia S., Rabino M., Pompilio G., Parini A., Lairez O., Gowran A, & Pizzinat N. (2023). IPSC derived cardiac fibroblasts of DMD patients show compromised actin microfilaments, metabolic shift and pro-fibrotic phenotype. Biology Direct, 18, 41.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!