Cellartis Human iPS Cell Line 12 (ChiPSC12, Y00285) was purchased from Takara Bio Inc. (Shiga, Japan). The cell line was created by reprogramming human skin fibroblasts using defective polycistronic retrovirus technology to deliver c-Myc, Oct-4, SOX2, and KLF4 [19 (link)–22 (link)]. The cell line origin has been confirmed by authentication using banded chromosome analysis. The last analysis was carried out a month before the start of the experiments. Mycoplasma testing was performed and all cell lines were used within 3 months of their acquisition. The cells were cultured using the Cellartis DEF-CS Culture System (Y30010, Takara Bio Inc.) including Matrigel-based culture system, according to the manufacturer’s instructions at 37 °C in 5% CO2. The cells were differentiated into the definitive endoderm using the Cellartis Definitive Endoderm Differentiation Kit (Y30030, Takara Bio Inc.).
Cellartis def cs culture system
Manufactured by Takara Bio
Sourced in Japan
The Cellartis DEF-CS Culture System is a laboratory equipment designed for the culturing and maintenance of human pluripotent stem cells. It provides a defined, feeder-free culture environment to support the growth and expansion of these cells.
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Lab products found in correlation
Essential 8 medium, by Thermo Fisher Scientific (1 mentions)
Hek293, by Health Science Research Resources Bank (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Cellartis hepatocyte differentiation kit, by Takara Bio (1 mentions)
Tryple select enzyme 1x, by Thermo Fisher Scientific (1 mentions)
Icell cardiomyocytes, by Fujifilm (1 mentions)
5 protocols using cellartis def cs culture system
1
Differentiation of iPSC into Definitive Endoderm
2
Maintenance of Human iPSC Lines
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The human iPS cell line 201B was provided by Dr. Shinya Yamanaka (Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, Kyoto, Japan). 201B cells were maintained in tissue culture plates coated with recombinant human truncated vitronectin (Invitrogen, Carlsbad, CA) in Essential 8 medium (Invitrogen) supplemented with 10 μM ROCK inhibitor Y27632 (Invitrogen). Cells were routinely passaged as small clumps using the EDTA method with the split ratio of 1:8 to 1:12 every 2 to 3 days after reaching 60% to 80% confluence [31 (link)]. The human iPSC line ChiPS17 was purchased from TaKaRa (Shiga, Japan) and cultured using Cellartis DEF-CS Culture System (TaKaRa) according to the manufacturer’s instructions. Human fibroblast BJ and cancer cell lines (K562, HeLa and HEK293) were purchased from the Health Science Research Resources Bank (Osaka, Japan) and maintained in RPMI1640 or DMEM medium (Sigma-Aldrich, St. Louis, MO) supplemented with 10% heat-inactivated fetal calf serum (FCS) (Sigma-Aldrich).
3
Hepatocyte Differentiation of 201B7 hiPSCs
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The 201B7 human iPS cell line44 (link) was acquired from the RIKEN BioResource Center (BRC) and was cultured in a Cellartis® DEF-CS™ Culture System (Takara Bio Inc., Shiga, Japan). For in vitro hepatocyte differentiation and DE differentiation, we used the Cellartis® Hepatocyte Differentiation Kit (Takara Bio Inc.) and the Cellartis® DE Differentiation Kit (Takara Bio Inc.), respectively, according to the manufacturers’ instructions. The culture conditions are shown in Fig. 1a .
4
iPSC Generation from Cord Blood
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Human iPSC cell OARSAi002-A was previously generated in our laboratory from the cord blood of a healthy non-Hispanic white male using self-replicative RNA reprogramming technology (15 (link)). Cells were maintained as a single-cell culture (no colony formation) on COAT-1 pre-coated 6-well tissue culture plates in the Cellartis DEF-CS Culture System (Takara Bio, Inc.) at 37˚C in a humidified incubator supplied with 5% CO2. For passaging, cells were dissociated into single cells with TrypLE Select Enzyme (1X) (Thermo Fisher Scientific, Inc.) and seeded at a density of 4-5x104 cells/cm2. The cell culture medium was changed daily until the cells reached a confluence of ~1.5-3.0x105 cells/cm2, which generally occurred 3-4 days post passage. The pluripotency of the iPSCs was routinely assessed using immunocytochemistry staining of major pluripotency protein markers, including octamer-binding transcription factor 4 (OCT4), SRY (sex determining region Y)-box (SOX)2, stage-specific embryonic antigen-4 (SSEA4) and T cell receptor α locus (TRA-1-60) using a Pluripotent Stem Cell 4-Marker Immunocytochemistry Kit (cat. no. A24881) according to the manufacturer's protocol (Thermo Fisher Scientific, Inc.).
5
Induced Pluripotent Stem Cell Cardiomyocytes
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For hiPSCs, Cellartis human iPS cell line 18 (ChiPSC18) cells were purchased from Takara Bio, Inc., and were maintained with the Cellartis DEF-CS culture system (Takara Bio, Inc.) according to the manufacturer's instructions. The cultured ChiPSC18 (passages 19-22) were frozen in Stem-Cellbanker (Zenoaq Resource Co. Ltd) and cryopreserved in liquid nitrogen. The frozen hiPSCs were thawed and used for experiments after two to four passages. For hiPSC-derived cardiomyocytes, iCell cardiomyocytes (#C1106, Lot#103700) were purchased from FUJIFILM Cellular Dynamics, Inc., and were used for experiments immediately after thawing using iCell cardiomyocyte plating medium (#M1001; FUJIFILM Cellular Dynamics, Inc.) according to the manufacturer's instructions.
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