Generation and Maintenance of Human iPSCs

The human iPSCs lines SFCi55 (parental, control) and SFCi55-iKLF1.2 were generated in house^{20 (link)}. The SFCi55 iPSC line was originally generated using fibroblasts obtained from blood group O Rhesus negative individuals by R Biomedical under REC 1/AL/0020 ethical approval and programmed to iPSCs using Yamanaka factors on episomal vectors. Both lines were confirmed to be pluripotent and have normal karyotype^{20 (link)}. All IPSC lines were routinely tested for mycoplasma and maintained in StemPro hESC SFM media (Gibco) with 20 ng/ml bFGF (R&D) (Maintenance media). Wells were pre-coated with CELLstart (Gibco) for 1 h before. Cells were passaged using the StemPro EZPassage tool (Thermo Fisher Scientific). Media change was performed every day and cells passaged at a ratio of 1:4 every 3–4 days, when cells reached 70% confluency.

Free full text: Click here

Lopez-Yrigoyen M., Yang C.T., Fidanza A., Cassetta L., Taylor A.H., McCahill A., Sellink E., von Lindern M., van den Akker E., Mountford J.C., Pollard J.W, & Forrester L.M. (2019). Genetic programming of macrophages generates an in vitro model for the human erythroid island niche. Nature Communications, 10, 881.

Publication 2019

bFGF CELLstart StemPro EZPassage tool

Cells Episomal Fibroblasts Hesc Human Ipsc Mycoplasma Parental Rhesus blood group Vectors

Corresponding Organization :

Other organizations : MRC Centre for Regenerative Medicine, University of Edinburgh, MRC Centre for Reproductive Health, Queen's Medical Centre, Scottish National Blood Transfusion Service, Sanquin, University of Amsterdam

Top 5 similar protocols

Variable analysis

independent variables

Generation of SFCi55-iKLF1.2 iPSC line from the parental SFCi55 iPSC line

dependent variables

Pluripotency of the iPSC lines
Karyotype of the iPSC lines

control variables

SFCi55 (parental, control) iPSC line
Maintenance of all iPSC lines in StemPro hESC SFM media with 20 ng/ml bFGF
Pre-coating of wells with CELLstart
Passaging of cells using the StemPro EZPassage tool
Media change every day and passaging at a ratio of 1:4 every 3-4 days when cells reached 70% confluency

positive controls

SFCi55 (parental, control) iPSC line

negative controls

Not mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!