Log In Sign up
The largest database of trusted experimental protocols

Recombinant human lif

Manufactured by Merck Group
Visit Supplier
Sourced in China

Recombinant human LIF is a laboratory reagent that functions as a cytokine, promoting the self-renewal and pluripotency of stem cells. It is produced using recombinant DNA technology.

Automatically generated - may contain errors

Lab products found in correlation

Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions) Glutamine, by Sartorius (2 mentions) Penicillin, by Sartorius (2 mentions) Streptomycin, by Sartorius (2 mentions) Non essential amino acids (neaa), by Sartorius (2 mentions) Pen strep solution, by Sartorius (2 mentions) Dulbecco modified eagle medium (dmem), by Sartorius (2 mentions) β mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Trypsin edta, by Thermo Fisher Scientific (2 mentions) Forskolin, by Merck Group (2 mentions) Tryple, by Thermo Fisher Scientific (2 mentions) Chir99021, by Selleck Chemicals (2 mentions) Neurobasal medium, by Thermo Fisher Scientific (2 mentions) Ca2 mg2, by Corning (2 mentions) Ack rbc lysis buffer, by Thermo Fisher Scientific (2 mentions) Neurobasal a medium, by Thermo Fisher Scientific (2 mentions) Dnase, by Worthington (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) C11995500bt, by Thermo Fisher Scientific (1 mentions)

13 protocols using recombinant human lif

1

Murine Embryonic Stem Cell Culture Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols
The G4 (C57BL/6Ncr x 129S6/SvEvTac) mouse hybrid (George et al., 2007 (link)) ESCs were obtained from Mount Sinai Hospital and were maintained on STO feeders in serum-containing media at 5% CO2 and 37°C. They were sub-cloned, and a line with normal karyotype was selected for further analysis. The cells were split onto gelatinized plates (10 cm, Corning) and expanded in serum-containing media or chemically defined media (standard 2i or alternative 2i) for at least three passages. Cells were harvested by trypsinization (0.05% trypsin/EDTA, GIBCO) for 10 min, at which point they reached 70%–80% confluence for single-cell capture.
The three media are as follows:

Serum-containing media: Knockout DMEM (GIBCO), 1X penicillin-streptomycin-glutamine (GIBCO), 1X non-essential amino acids (GIBCO), 100 U/ml recombinant human leukemia inhibitory factor (Millipore), 15% fetal bovine serum (HyClone), 0.1mM β-mercaptoethanol (Sigma).

Standard 2i media: N2B27 basal media (NDiff 227, StemCells), 100 U/ml recombinant human LIF (Millipore), 1 μM PD0325901 (Stemgent), 3 μM CHIR99021 (Stemgent).

Alternative 2i media: N2B27 basal media (NDiff 227, StemCells), 100 U/ml recombinant human LIF (Millipore), 1 μM CGP77675 (Sigma), 3 μM CHIR99021 (Stemgent).

Kolodziejczyk A.A., Kim J.K., Tsang J.C., Ilicic T., Henriksson J., Natarajan K.N., Tuck A.C., Gao X., Bühler M., Liu P., Marioni J.C, & Teichmann S.A. (2015). Single Cell RNA-Sequencing of Pluripotent States Unlocks Modular Transcriptional Variation. Cell Stem Cell, 17(4), 471-485.
+ Open protocol
+ Expand
2

Chordoma Cell Lines Characterization

Check if the same lab product or an alternative is used in the 5 most similar protocols
Chordoma cell lines U-CH120 (link) and MUG-Chor121 (link), available from the ATCC Bioresource Center, were provided by the Chordoma Foundation (Durham, NC, USA). Cells were cultured in culture flasks or well plates coated with 0.1% gelatin (Cat. No. G1890-100G; Sigma-Aldrich) and IMDM/RPMI (4:1) containing 10% fetal bovine serum and 1% antibiotics. Cell lines were checked routinely for molecular markers. Polymerase chain reaction (PCR)-based mycoplasma screening and Sanger sequencing were used for short tandem repeat to verify retaining of chordoma character. Recombinant human LIF was obtained from Millipore (Cat. No. LIF 1050) and administered to cells in medium in a 100 ng/ml concentration.
Gulluoglu S., Sahin M., Tuysuz E.C., Yaltirik C.K., Kuskucu A., Ozkan F., Sahin F., Ture U, & Bayrak O.F. (2017). Leukemia Inhibitory Factor Promotes Aggressiveness of Chordoma. Oncology Research, 25(7), 1177-1188.
+ Open protocol
+ Expand
3

Cell Culture and Immunoblot Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols
HepG2, 293T and PC3 cells were obtained from the Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS, Shanghai). Cells were cultured in high glucose DMEM (C11995500BT, Life Technologies) containing 10% fetal bovine serum (FBS) (10099–141, Life Technologies) with 100 units/ml penicillin and 10 μg/ml streptomycin (15140–122, Life Technologies) at 37°C in a 5% CO2 atmosphere.
The polyclonal antibody against STAT3 C-20 (sc-482) and monoclonal antibodies for Myc (sc-40) and pY20 (sc-508) were from Santa Cruz Biotechnology, Inc. The GFP monoclonal antibody (11814460001) was from Roche and the polyclonal antibody for acetyl-lysine (#9441s) was from Cell Signaling Technology. The secondary antibodies, including goat anti-rabbit RDye® 680RD (926–68071) and goat anti-mouse RDye® 800CW (926–32210), were from LICOR. Anti-pY45-STAT3 and anti-acetyl-K78-STAT3 polyclonal antibodies were prepared by AB-land, Inc. (Hangzhou, China).
Recombinant human IL-6 was from Life Technologies, LPS was from Sigma and recombinant human LIF was from Millipore. The dual-luciferase reporter assay system kit (E1910) was obtained from Promega.
Xu L., Ji J.J., Le W., Xu Y.S., Dou D., Pan J., Jiao Y., Zhong T., Wu D., Wang Y., Wen C., Xie G.Q., Yao F., Zhao H., Fan Y.S, & Chin Y.E. (2015). The STAT3 HIES mutation is a gain-of-function mutation that activates genes via AGG-element carrying promoters. Nucleic Acids Research, 43(18), 8898-8912.
+ Open protocol
+ Expand
4

Cytokine Signaling Pathway Modulation

Check if the same lab product or an alternative is used in the 5 most similar protocols
TGFβ1 was purchased from Peprotech (#100-21, Peprotech, Rocky Hill, NJ) and was used at 2 ng/ml; recombinant human GCSF (#300-23) and IL-6 (#200-06) were purchased from Peprotech and were used at 10ng/mL, recombinant human LIF was purchased from Millipore (#LIF1005, Millipore, Billerica, MA), and was used at a concentration of 2 ng/ml. Recombinant TNF alpha was produced in E Coli and purified under native conditions using an N-terminal 6-his tag. ICAM-1 neutralizing antibody (#BBA3, R&D, Minneapolis, MN) was used at 10 μg/ml. The following inhibitors were used in this study: Ruxolitinib (#1598, Axon medchem, Groningen, The netherlands) at 10μM, CYT387 (#S2219, Selleckchem, Huston, TX) at 10μM Y27632 (#1254, Tocris bioscience, Ellisville, MO) at 10μM, Blebbistatin (#B0560, Sigma, Saint Louis, MO) at 10μM, SU6656 (#572635, CalbioChem, Los Angeles, CA) at 10μM and Verteprofin (#SML0534, Sigma, Saint Louis, MO) at 4μg/mL.
Bonan S., Albrengues J., Grasset E., Kuzet S.E., Nottet N., Bourget I., Bertero T., Mari B., Meneguzzi G, & Gaggioli C. (2016). Membrane-bound ICAM-1 contributes to the onset of proinvasive tumor stroma by controlling acto-myosin contractility in carcinoma-associated fibroblasts. Oncotarget, 8(1), 1304-1320.
+ Open protocol
+ Expand
5

Culturing Cell Lines and Organoids

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
All cell lines were cultured in a humidified incubator at 37° C and 5% C02 in medium supplemented with 10% fetal calf serum (FCS) and Pen/Strep solution (Biological industries). The human colon carcinoma cell line SW480 was maintained in DMEM (Biological industries). The human colon cancer isogenic RKO cell lines were kindly provided by Prof. Bert Vogelstein (The Johns Hopkins University, MD, USA) and maintained in McCoy’s 5A medium supplemented with 2mM L-glutamine. The non-small lung carcinoma cell line, NCI-H1299, was maintained in RPMI-1640. iPSCs were generated as previously described [31 (link)] and maintained on irradiated MEFs in ES medium [DMEM (Biological Industries) containing 15% FCS, 5 mg recombinant human LIF (Millipore), 1 mM glutamine (Biological Industries), 1% nonessential amino acids (Biological Industries), 0.1 mM β-mercaPtoethanol (Invitrogen), 60μg/ml penicillin, and 100 μg/ml streptomycin (Biological Industries)]. Intestinal organoids were generated as previously described [15 (link)]. SK-BR-3 breast cancer cells were maintained in RPMI-1640 medium. Mycoplasma test is being performed routinely.
Solomon H., Dinowitz N., Pateras L.S., Cooks T., Shetzer Y., Molchadsky A., Charni M., Rabani S., Koifman G., Tarcic O., Porat Z., Kogan-Sakin I., Goldfinger N., Oren M., Harris C.C., Gorgoulis V.G, & Rotter V. (2018). Mutant p53 gain of function underlies high expression levels of colorectal cancer stem cells markers. Oncogene, 37(12), 1669-1684.
+ Open protocol
+ Expand
6

Isolation and Culture of Murine Embryonic Stem Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
SCNT blastocysts with a well-developed ICM on Day 6.5 or Day 7 were used for isolating ICM as described previously7 (link). Zonae pellucidae of these blastocysts were removed with 1% pronase (Sigma), and were manually split into two portions. Part of containing the ICM was pressed onto mouse embryonic fibroblast (MEF) feeder layer inactivated with mitomycin C (10 μg/ml; Sigma), and allowed to form primary embryo outgrowths. The ICM was cultured in a single well of 4-well or 24-well plate (Nunc) containing 1 ml medium consisting of basic ES medium (KnockOut-Dulbecco modified Eagle medium supplemented with 2 mM Glutamine, 1% MEM nonessential amino acids, 20 ng/ml human recombinant basic fibroblasts growth factor, 20 ng/ml recombinant human LIF (Millipore), and 0.1 mM β-mercaptoethanol) supplemented with 15% ES cell qualified FBS (Hyclone) under 37 °C, 5% CO2. After 6 to 7 days of culture, compact ICM outgrowth was removed by a glass pipette and transferred onto new feeder layers with basic ES medium supplemented with 10% FBS and 10% KnockOut-Serum Replacement (KSR). The medium was changed every two days. About one week, the colonies were broken into small pieces by mouth pipette, gently, and then transferred onto new feeder layer with basic ES medium supplemented with 20% KSR.
Wu X., Song M., Yang X., Liu X., Liu K., Jiao C., Wang J., Bai C., Su G., Liu X, & Li G. (2016). Establishment of bovine embryonic stem cells after knockdown of CDX2. Scientific Reports, 6, 28343.
+ Open protocol
+ Expand
7

Culturing Cell Lines and Organoids

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
All cell lines were cultured in a humidified incubator at 37° C and 5% C02 in medium supplemented with 10% fetal calf serum (FCS) and Pen/Strep solution (Biological industries). The human colon carcinoma cell line SW480 was maintained in DMEM (Biological industries). The human colon cancer isogenic RKO cell lines were kindly provided by Prof. Bert Vogelstein (The Johns Hopkins University, MD, USA) and maintained in McCoy’s 5A medium supplemented with 2mM L-glutamine. The non-small lung carcinoma cell line, NCI-H1299, was maintained in RPMI-1640. iPSCs were generated as previously described [31 (link)] and maintained on irradiated MEFs in ES medium [DMEM (Biological Industries) containing 15% FCS, 5 mg recombinant human LIF (Millipore), 1 mM glutamine (Biological Industries), 1% nonessential amino acids (Biological Industries), 0.1 mM β-mercaPtoethanol (Invitrogen), 60μg/ml penicillin, and 100 μg/ml streptomycin (Biological Industries)]. Intestinal organoids were generated as previously described [15 (link)]. SK-BR-3 breast cancer cells were maintained in RPMI-1640 medium. Mycoplasma test is being performed routinely.
Solomon H., Dinowitz N., Pateras L.S., Cooks T., Shetzer Y., Molchadsky A., Charni M., Rabani S., Koifman G., Tarcic O., Porat Z., Kogan-Sakin I., Goldfinger N., Oren M., Harris C.C., Gorgoulis V.G, & Rotter V. (2018). Mutant p53 gain of function underlies high expression levels of colorectal cancer stem cells markers. Oncogene, 37(12), 1669-1684.
+ Open protocol
+ Expand
8

Conversion of Human iPSCs to Naive State

Check if the same lab product or an alternative is used in the 5 most similar protocols
Human primed iPSCs were treated with 10 μM Rho-associated kinase (ROCK) inhibitor Y27632 (Abcam) overnight and then trypsinized into single cells with 0.25% trypsin/EDTA (Life Technologies) for 10 min at 37°C. The single cells were plated on MEF feeders (5–6 × 105/cm2) in naivetropic iPSC medium (50% DMEM/F12 and 50% Neurobasal, with N2, B27, 1 mM glutamine, 1% NEAA, 0.1 mM β-mercaptoethanol, penicillin–streptomycin (all from Life Technologies), 20 ng/mL recombinant human LIF (Millipore), 5 mg/mL bovine serum albumin (BSA) (Sigma), 2 μg/mL doxycycline (Sigma), 1 μM PD0325901 (EMD Millipore), and 3 μM CHIR99021 (Stemgent) and cultured for 7–10 days. The small, bright, compact cell clumps in the culture were picked manually and trypsinized into single cells, which were replated on fresh MEF feeders. After several passages, mouse ESC-like dome-shaped colonies were trypsinized to single cells and passaged every 2–3 days. Naivetropic iPSCs were derived and cultured in an incubator with 5% O2.
Hu Z., Pu J., Jiang H., Zhong P., Qiu J., Li F., Wang X., Zhang B., Yan Z, & Feng J. (2015). Generation of Naivetropic Induced Pluripotent Stem Cells from Parkinson's Disease Patients for High-Efficiency Genetic Manipulation and Disease Modeling. Stem Cells and Development, 24(21), 2591-2604.
+ Open protocol
+ Expand
9

Derivation and Maintenance of Human Naive PSCs

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
To derive human naive PSCs, primed cells were switched to naive media based on N2B27 or mTeSR. N2B27 medium consisted of a 1:1 mixture of DMEM/F12 and Neurobasal medium, with 1% N2 supplement, 2% B27 supplement, 1 mM L-glutamax, 0.1 mM beta-mercaptoethanol, 1× non-essential amino acids, and 1× penicillin-streptomycin (all from Invitrogen). The mTeSR1 medium was purchased from STEMCELL. N2B27 and mTeSR base media were supplemented with 2iFL: PD0325901 (Stemgent, 0.5 μM), CHIR99021 (Selleck Chemicals, 3 μM), Forskolin (Sigma, 10 μM) and recombinant human LIF(EMD Millipore, 10 ng/ml). LPA (Sigma, 10 μM) was added to naive media when indicated. The ROCK inhibitor Y-27632 (Stemgent, 5 μM) was used for the first three passages. Human naive PSCs were maintained on irradiated MEFs. To passage naive PSCs, cells were briefly washed with PBS, dissociated to single cells using TrypLE (Invitrogen), centrifuged in fibroblast medium (DMEM with 10% fetal bovine serum, 1 mM glutamax, 1× non-essential amino acids, 1× sodium pyruvate, 1× penicillin/streptomycin, and 0.06 mM β-mercaptoethanol, all from Invitrogen), and seeded onto irradiated MEFs in naive medium. The 5i/L/A medium was prepared as described earlier (Theunissen et al., 2014 (link)).
Qin H., Hejna M., Liu Y., Percharde M., Wossidlo M., Blouin L., Durruthy-Durruthy J., Wong P., Qi Z., Yu J., Qi L.S., Sebastiano V., Song J.S, & Ramalho-Santos M. (2016). YAP Induces Human Naive Pluripotency. Cell reports, 14(10), 2301-2312.
+ Open protocol
+ Expand
10

Derivation and Maintenance of Human Naive PSCs

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
To derive human naive PSCs, primed cells were switched to naive media based on N2B27 or mTeSR. N2B27 medium consisted of a 1:1 mixture of DMEM/F12 and Neurobasal medium, with 1% N2 supplement, 2% B27 supplement, 1 mM L-glutamax, 0.1 mM beta-mercaptoethanol, 1× non-essential amino acids, and 1× penicillin-streptomycin (all from Invitrogen). The mTeSR1 medium was purchased from STEMCELL. N2B27 and mTeSR base media were supplemented with 2iFL: PD0325901 (Stemgent, 0.5 μM), CHIR99021 (Selleck Chemicals, 3 μM), Forskolin (Sigma, 10 μM) and recombinant human LIF(EMD Millipore, 10 ng/ml). LPA (Sigma, 10 μM) was added to naive media when indicated. The ROCK inhibitor Y-27632 (Stemgent, 5 μM) was used for the first three passages. Human naive PSCs were maintained on irradiated MEFs. To passage naive PSCs, cells were briefly washed with PBS, dissociated to single cells using TrypLE (Invitrogen), centrifuged in fibroblast medium (DMEM with 10% fetal bovine serum, 1 mM glutamax, 1× non-essential amino acids, 1× sodium pyruvate, 1× penicillin/streptomycin, and 0.06 mM β-mercaptoethanol, all from Invitrogen), and seeded onto irradiated MEFs in naive medium. The 5i/L/A medium was prepared as described earlier (Theunissen et al., 2014 (link)).
Qin H., Hejna M., Liu Y., Percharde M., Wossidlo M., Blouin L., Durruthy-Durruthy J., Wong P., Qi Z., Yu J., Qi L.S., Sebastiano V., Song J.S, & Ramalho-Santos M. (2016). YAP Induces Human Naive Pluripotency. Cell reports, 14(10), 2301-2312.
+ 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!