====== Cell lines ====== In FG Kaessmann we use only induced pluripotent stem cells (iPSCs). Here are the cell lines we have: ===== Human ===== === SCTi === full name: SCTi003-A - a cell line bought by us from STEMCELL Technologies\\ in FG kaessmann?: yes\\ biosafety level: S1\\ GMO: yes (because ->)\\ reprogramming method: plasmid or virus\\ cell bank created: yes\\ cultured in: mTESR plus, no antibiotics\\ cultured on: Matrigel\\ passaged with: ReLeSR, 5 min 37C, no anti-apoptotic after\\ passaged in, ratio: clumps, 1:20-1:30\\ individual: female\\ cell line of origin: PBMC\\ original culturing [[https://www.stemcell.com/products/healthy-control-human-ipsc-line-female-scti003-a.html|protocol]]\\ cell line reference/registration:[[https://hpscreg.eu/browse/provider/484|hPSCreg database entry]] note: so far can't be used for further experiments. StemCell Technologies have to provide the info on reprogramming procedure. === IDP === IDP52555 (sometimes marked as ZNF492-WT or ZNF) - a cell line, shared with us by Michael Heide from Göttingen\\ in FG kaessmann?: yes\\ biosafety level: S1\\ GMO: yes (because ->)\\ reprogramming method: virus, [[https://www.thermofisher.com/order/catalog/product/de/en/A16517|reprogramming kit]], similar to the [[https://www.biorxiv.org/content/10.1101/2022.12.07.519435v1.abstract|protocol]] \\ cell bank created: yes\\ cultured in: mTESR plus, no antibiotics\\ cultured on: Geltrex or Matrigel\\ passaged with: ReLeSR, 5 min 37C, no anti-apoptotic after\\ passaged in, ratio: clumps, 1:6-1:10\\ individual: female\\ cell line of origin: fibroblasts\\ original culturing [[wetlab:cell_culture:cell_lines_list:znf_protocol|protocol]]\\ cell line reference/registration: [[https://hpscreg.eu/cell-line/CRTDi011-A|hPSCreg database entry]] === rep771 === Repro 771-3G - a cell lines, shared with us by Lena Kutscher from DKFZ\\ in FG kaessmann?: yes\\ biosafety level: S1\\ GMO: no\\ reprogramming method: mRNA, [[https://www.reprocell.com/product-catalog/rna-reprogramming/stemrna-3rd-gen-reprogramming-kit|StemRNA™ 3rd Gen Reprogramming Kit]]\\ cell bank created: not yet\\ cultured in: mTESR plus, no antibiotics\\ cultured on: Matrigel\\ passaged with: ReLeSR, 5 min 37C, no anti-apoptotic after\\ passaged in, ratio: clumps, 1:10-1:15\\ individual: male\\ cell line of origin: endothelial progenitor cells (from blood)\\ cell line reference/registration:[[https://www.reprocell.com/product-catalog/induced-pluripotent-stem-cells/stemrna-human-ipsc-771-3g| ReproCell]] ===== Chimp ===== === JoC === JoC - a cell lines, shared with us by Svante Pääbo from MPI, Leipzig\\ in FG kaessmann?: yes\\ biosafety level: S2 (not tested for HIV)\\ GMO: no\\ reprogramming method: plasmids from [[https://doi.org/10.1002/stem.1293|Okita et al, 2013]] \\ cell bank created: not yet\\ cultured in: mTESR plus, no antibiotics\\ cultured on: Matrigel\\ passaged with: will add later\\ passaged in, ratio: will add later\\ individual: ?\\ cell line of origin: primary lymphocytes (from blood)\\ original culturing [[wetlab:cell_culture:cell_maintenance:liepzig_primate|Leipzig cell culture protocol]]\\ cell line reference/registration: [[https://doi.org/10.1038/s41586-019-1654-9|Kanton et al, 2019]] === Sandra === Sandra - a cell lines, shared with us by Svante Pääbo from MPI, Leipzig\\ in FG kaessmann?: yes\\ biosafety level: S1 \\ GMO: no\\ reprogramming method: plasmids from [[https://doi.org/10.1002/stem.1293|Okita et al, 2013]] \\ cell bank created: yes\\ cultured in: mTESR plus, no antibiotics\\ cultured on: Matrigel\\ passaged with: ReLeSR, 5 min 37C, no anti-apoptotic after\\ passaged in, ratio: clumps, 1:6-1:10\\ individual: ?\\ cell line of origin: leukocytes (from blood)\\ original culturing [[wetlab:cell_culture:cell_maintenance:liepzig_primate|Leipzig cell culture protocol]]\\ cell line reference/registration: [[https://doi.org/10.7554/eLife.18683|Mora-Bermúdez et al, 2016]] === Karlchen === Karlchen - a cell lines, shared with us by Sebastian Diecke from MDC, Berlin\\ in FG kaessmann?: yes\\ biosafety level: S2 \\ GMO: yes\\ reprogramming method: virus, [[https://www.thermofisher.com/order/catalog/product/de/en/A16517|reprogramming kit]] \\ cell bank created: not yet\\ cultured in: E8 (mTESR on the vial), no antibiotics\\ cultured on: Matrigel\\ passaged with: EDTA/versene, X, X\\ passaged in, ratio: clumps, X\\ individual: male\\ cell line of origin: fibroblasts\\ cell line reference/registration: [[https://doi.org/10.1016/j.devcel.2020.07.006|Wilson et al, 2020]] ===== Gorilla ===== === Assumbo 1 === Assumbo 1 PBMC - a cell lines, shared with us by Sebastian Diecke from MDC, Berlin\\ in FG kaessmann?: yes\\ biosafety level: S2 \\ GMO: yes\\ reprogramming method: virus, [[https://www.thermofisher.com/order/catalog/product/de/en/A16517|reprogramming kit]] \\ cell bank created: not yet\\ cultured in: E8 (mTESR on the vial), no antibiotics\\ cultured on: Matrigel\\ passaged with: EDTA/versene, X, X\\ passaged in, ratio: clumps, X\\ individual: number 1, male, same as Assumbo 2\\ cell line of origin: PBMC cell line reference/registration: [[https://doi.org/10.1016/j.devcel.2020.07.006|Wilson et al, 2020]]\\ note: not QCed by MDC === Assumbo 2 === Assumbo 2 fib - a cell lines, shared with us by Sebastian Diecke from MDC, Berlin\\ in FG kaessmann?: yes\\ biosafety level: S2 \\ GMO: yes\\ reprogramming method: virus, [[https://www.thermofisher.com/order/catalog/product/de/en/A16517|reprogramming kit]] \\ cell bank created: not yet\\ cultured in: E8 (mTESR on the vial), no antibiotics\\ cultured on: Matrigel\\ passaged with: EDTA/versene, X, X\\ passaged in, ratio: clumps, X\\ individual: number 1, male, same as Assumbo 1\\ cell line of origin: fibroblasts\\ cell line reference/registration: [[https://doi.org/10.1016/j.devcel.2020.07.006|Wilson et al, 2020]]\\ note: QC performed by MDC ===== Rhesus macaque ===== === Rh33.1 === a cell line, shared with us by Rüdiger Behr from Göttingen\\ in FG kaessmann?: yes\\ biosafety level: S1\\ GMO: yes\\ reprogramming method: plasmids transfection, cassette integrated, clone 33.1 ([[https://www.mdpi.com/2073-4409/9/6/1349|publication]])\\ cell bank created: yes\\ cultured in: iPSC Brew, 1uM IWR1, 0.5uM CHIR, no antibiotics\\ cultured on: Geltrex\\ passaged with: versene, 3 min RT, add new versene, 3 min RT, add anti-apoptotic after passaging\\ passaged in, ratio: clumps, 1:10-1:15\\ individual: 002468 (DPZ number), male\\ original culturing [[wetlab:cell_culture:cell_lines_list:rh_protocol|protocol]]\\ cell line reference/registration: [[https://doi.org/10.3390/cells9061349|Stauske et al, 2020]] === Rh24.4 === A cell line, shared with us by Rüdiger Behr from Göttingen\\ in FG kaessmann?: yes\\ biosafety level: S1\\ GMO: yes\\ reprogramming method: plasmids transfection, no cassette integration, clone 24.4 ([[https://www.mdpi.com/2073-4409/9/6/1349|publication]])\\ cell bank created: yes\\ cultured in: iPSC Brew, 1uM IWR1, 0.5uM CHIR, no antibiotics\\ cultured on: Geltrex\\ passaged with: versene, 3 min RT, add new versene, 3 min RT, add anti-apoptotic after passaging\\ passaged in, ratio: clumps, 1:10-1:15\\ individual: 002616 (DPZ number), female\\ original culturing [[wetlab:cell_culture:cell_lines_list:rh_protocol|protocol]]\\ cell line reference/registration: [[https://doi.org/10.3390/cells9061349|Stauske et al, 2020]] ===== Marmoset ===== === CJ lines === cj_080619_4, cj_160919_5, cj_160919_6 - 3 cell lines, shared with us by Rüdiger Behr from Göttingen\\ in FG kaessmann?: yes\\ biosafety level: S1\\ GMO: no\\ reprogramming method: mRNA, clones 1-3 ([[https://www.mdpi.com/2073-4409/9/11/2422|publication]])\\ cell bank created: no (waiting for hypoxia incubator)\\ cultured in: iPSC Brew, 1uM IWR1, 0.5uM CHIR, no antibiotics, hypoxia\\ cultured on: Geltrex\\ passaged with: versene\\ passaged in, ratio: clumps, 1:10-1:15\\ individual: 018008; 017811; 017812 (DPZ number), gender ? \\ original culturing will add later\\ cell line registration: NA