Capan‐1, a H type‐3‐positive human pancreatic cancer cell line,8 was obtained from American Type Culture Collection (ATCC; Cat# HTB‐79, CVCL_0237). SUIT‐2, a H type‐3‐negative human pancreatic cancer cell line, was obtained from Japanese Collection of Research Bioresources (JCRB) Cell Bank (Cat# JCRB1094, CVCL_3172). All human cell lines have been authenticated using STR profiling within the last 3 years. All experiments were performed with mycoplasma‐free cells. Capan‐1 cells were cultured in Iscove's modified Dulbecco's medium (FUJIFILM Wako Pure Chemical Corporation, Cat# 098‐06465) supplemented with 20% fetal bovine serum (FBS; Thermo Fisher Scientific, Cat# 10270‐106) and containing 1% Penicillin‐Streptomycin (FUJIFILM Wako Pure Chemical Corporation, Cat# 168‐23191). SUIT‐2 cells were cultured in Dulbecco's modified Eagle's medium (FUJIFILM Wako Pure Chemical Corporation, Cat# 044‐29765) supplemented with 10% FBS and containing 1% Penicillin‐Streptomycin. All cells were maintained at 37°C in a humidified incubator in 5% CO2.
Iscove s modified dulbecco s medium
Manufactured by Fujifilm
Iscove's modified Dulbecco's medium is a cell culture medium used to support the growth and maintenance of various cell types. It is a chemically defined, serum-free medium that provides the necessary nutrients and supplements for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Capan 1, by Fujifilm (3 mentions)
Suit 2, by Fujifilm (3 mentions)
Dulbecco s modified eagle s medium, by Fujifilm (3 mentions)
Penicillin streptomycin, by Fujifilm (3 mentions)
Suit 2, by Japanese Collection of Research Bioresources Cell Bank (2 mentions)
Capan 1, by American Type Culture Collection (2 mentions)
Suit 2 cells, by Japanese Collection of Research Bioresources Cell Bank (1 mentions)
Cas9 smart nuclease all in one vector, by System Biosciences (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
4 protocols using iscove s modified dulbecco s medium
1
Culturing Pancreatic Cancer Cell Lines
2
Culturing Pancreatic Cancer Cell Lines
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The human pancreatic cancer cell line Capan‐1 was purchased from the American Type Tissue Culture Collection (ATCC), and SUIT‐2 cells were obtained from the Japanese Collection of Research Bioresources Cell Bank. The culture media and conditions were used as previously described.4 (link) Capan‐1 cells were cultured in Iscove's modified Dulbecco's medium (FUJIFILM Wako Pure Chemical Co) supplemented with 20% fetal bovine serum (Thermo Fisher Scientific) and 1% penicillin‐streptomycin (FUJIFILM Wako Pure Chemical Co). SUIT‐2 cells were cultured in Dulbecco's modified Eagle's medium (FUJIFILM Wako Pure Chemical Co) supplemented with 10% FBS and 1% penicillin‐streptomycin (FUJIFILM Wako Pure Chemical Co)
3
CRISPR/Cas9-Mediated FKTN Knockout in Cell Lines
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The CRISPR/Cas9 targeting sequence for FKTN was GAGTAGAATCAATAAGAACG. The oligonucleotides for this sequence were inserted into the Cas9 Smart nuclease all-in-one vector (System Biosciences, Mountain View, CA) with an additional 8-base sequence at the 5′ terminus. The vector was transfected into HEK293 (American Type Culture Collection, Manassas, VA, USA; CRL-1573) or HAP1 cells (Haplogen, Vienna, Austria; C631). IIH6-negative cells were sorted by fluorescence-activated cell sorting (MoFlo, Beckman Coulter, Brea, CA). HEK293 cells were cultured in DMEM supplemented with 10% FCS and penicillin/streptomycin. HAP1 cells were cultured in Iscove’s modified Dulbecco’s medium (Wako Pure Chemical Industries) supplemented with 10% FCS and penicillin/streptomycin. Each cell clone was verified for IIH6 reactivity by Western blot analysis and DNA sequencing. The mutations in each clone were as follows: (HEK293 cell is a triploid) HEK FKTN1o-2 (27-bp deletion, 1-bp deletion, and 1-bp insertion in exon 2, causing frameshifts) and HAP FKTN5 (34-bp deletion in exon 2, causing a frameshift).
4
Cultivation of Pancreatic Cancer Cell Lines
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Capan‐1, an H‐type‐3‐positive human pancreatic cancer cell line, was obtained from American Type Culture Collection (ATCC; Cat# HTB‐79). SUIT‐2, an H‐type‐3‐negative human pancreatic cancer cell line, was obtained from the Japanese Collection of Research Bioresources (JCRB) Cell Bank (Cat# JCRB1094, CVCL_3172). Cells were routinely tested for Mycoplasma and authenticated by the JCRB Cell Bank using short tandem repeat analysis. Capan‐1 cells were cultured in Iscove's Modified Dulbecco's Medium (FUJIFILM Wako Pure Chemical Corporation, Cat# 098‐06465) supplemented with 20% fetal bovine serum (FBS; Thermo Fisher Scientific, Cat# 10270‐106) and 1% penicillin–streptomycin (FUJIFILM Wako Pure Chemical Corporation, Cat# 168‐23191). SUIT‐2 cells were cultured in Dulbecco's modified Eagle's medium (FUJIFILM Wako Pure Chemical Corporation, Cat# 044‐29765) supplemented with 10% FBS and containing 1% penicillin‐streptomycin. Cells were maintained at 37°C in a humidified incubator in 5% CO2.
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