The human skin fibroblast cell line NB1RGB was obtained from the RIKEN BioResource Center (Ibaraki, Japan). The NB1RGB was maintained in Dulbecco’s modified Eagle’s medium (DMEM; Nacalai Tesque, Japan) supplemented with 10% fetal bovine serum (FBS; Hyclone, GE Healthcare, Chicago, IL, USA) and penicillin/streptomycin (Nacalai Tesque) at 37°C under 5% CO2 conditions. The iPSCs derived from NB1RGB were cultured by feeder-free methods using precoating with iMatrix511 (Nippi, Japan) and maintained in NutriStem™ XF/FF (Stemgent, Beltsville, MD, USA) culture medium at 37°C under 5% CO2 conditions. The hiPSC cell line 201B7 was obtained from the RIKEN BioResource Center and adapted from a fed-batch culture to a feeder-free culture using iMatrix and NutriStem™ XF/FF. NPCs derived from iPSCs were maintained in a PSC neural basal medium (Gibco, Thermo Fisher Scientific, Waltham, MA, USA). To prevent apoptosis, the iPSC and NPC culture media were supplemented with the Y27632 ROCK inhibitor (WAKO Pure Chemical Industries, Tokyo, Japan) during passage; on the subsequent day, the media were replaced again but without the Y27632 ROCK inhibitor.
Y27632 rock inhibitor
Manufactured by Fujifilm
Sourced in Japan
Y27632 ROCK inhibitor is a chemical compound that is used as a selective and potent inhibitor of Rho-associated protein kinase (ROCK). ROCK is an enzyme that plays a role in various cellular processes, including cell migration, proliferation, and apoptosis. Y27632 ROCK inhibitor is commonly used in cell culture and research applications to study the effects of ROCK inhibition on cellular behavior.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Nacalai Tesque (2 mentions)
Nb1rgb, by Nacalai Tesque (2 mentions)
Nb1rgb, by RIKEN BioResource Center (2 mentions)
Fetal bovine serum (fbs), by GE Healthcare (1 mentions)
Dulbecco s modified eagle s medium dmem, by Nacalai Tesque (1 mentions)
Imatrix 511, by Nippi (1 mentions)
Cell proliferation dye efluor 670, by Thermo Fisher Scientific (1 mentions)
Growth factor reduced matrigel, by BD (1 mentions)
Neural basal medium, by Thermo Fisher Scientific (1 mentions)
Psc neural induction medium, by Thermo Fisher Scientific (1 mentions)
Imatrix 511 silk, by Nippi (1 mentions)
3 protocols using y27632 rock inhibitor
1
Feeder-free Derivation and Maintenance of iPSCs and NPCs
2
Murine Lung Epithelial-Fibroblast Co-Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Epithelial cells (5 × 103) derived from E13.5, E15.5, E18.5, P14, or P56 murine lungs and fibroblasts (1 × 105) derived from P56 murine lungs were sorted into 500 µL of MTEC/Plus medium40 (link), resuspended in MTEC/Plus medium, and mixed 1:1 with growth factor–reduced Matrigel (BD Biosciences). The resulting 90 µl mixture of was placed in a 24-well 0.4-μm Transwell clear insert (Falcon; BD Biosciences)14 (link). Next, 500 µl MTEC/Plus medium was added to the lower chamber, and MTEC/Plus medium was changed every other day14 (link). The Y-27632 ROCK inhibitor (10 μM; Wako Pure Chemical Industries) was added to the medium for the first 2 days. Images were acquired after 14 days of culture. Colony sizes and numbers were quantified using ImageJ version 1.8 (NIH, Bethesda, MD; http://imagej.nih.gov/ij ). Colony forming efficiency was defined as the number of colonies formed divided by 5,000 (number of cells plated). For proliferation dye assay, sorted epithelial cells were labeled with Cell Proliferation Dye eFluor 670 (Thermo Fisher Scientific), and 5 × 103 cells were used for co-culture with 1 × 105 fibroblasts derived from P56 murine lungs.
3
Induced Pluripotent Stem Cell Derivation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human skin fibroblasts NB1RGB [20–22 (link)] and human iPSCs 201B7 [23 (link)] were obtained from RIKEN Bio Resource Center. NB1RGB was maintained with Dulbecco’s Modified Eagle Medium and supplemented with 10% FBS and penicillin/streptomycin (Nacalai Tesque, Japan). iPSCs C2 were derived from NB1RGB by messenger RNA (mRNA) integration-free methods using the Stemgent® StemRNATM-NM Reprogramming Kit for Reprogramming Adult and Neonatal Human Fibroblasts (Stemgent, USA) as previously reported [11 (link), 24 (link)]. iPSCs were maintained as feeder-free culture with the NutriStem™ XF/FF medium (ReproCELL, Japan) and iMatrix511 silk (Nippi, Japan). NPCs were derived from iPSCs C2 using the PSC neural induction medium (Gibco, Thermo Fisher Scientific, USA) and maintained with the neural basal medium (Gibco, Thermo Fisher Scientific) and iMatrix 511 silk according to the manufacturer’s protocols with small modifications. iPSCs and NPCs culture media were supplemented with the Y27632 ROCK inhibitor (WAKO Pure Chemical Industries, Japan) during passage; on the subsequent day, the media were replaced with fresh culture media without the ROCK inhibitor.
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
Revolutionizing how scientists
search and build protocols!