NEUROD2 nVenus/nVenus iPSCs
HMGUi001-A-42
General
Cell Line |
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| hPSCreg name | HMGUi001-A-42 |
| Cite as: | HMGUi001-A-42 |
| Alternative name(s) |
NEUROD2 nVenus/nVenus iPSCs
|
| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines |
HMGUi001-A (XM001, BIHi043-A) HMGUi001-A-8 (C-PEP-mCherry-hiPSC) HMGUi001-A-43 (hINS-T2A-H2B-Cherry (-/-), hiPSC-INS-T2A-H2B-Cherry reporter, INSCherry/Cherry) HMGUi001-A-5 (ΔINK4 T2D risk region hiPSC) HMGUi001-A-1 (hINS-T2A-H2B-Cherry (+/-), HMGUi001-A-1) HMGUi001-A-4 (hiPSC-ARX-T2A-H2B-CFP-Flag) HMGUi001-A-22 (NCS1-KO Clone 19) HMGUi001-A-46 (ARX-CFP/PAX4-mCherry, ARX-T2A-H2B-CFP/H2B-mCherry-RGSHis-T2A-PAX4, ARXnCFP/nCFP/PAX4mCherry/mCherry) |
| Last update | 31st August 2023 |
| Notes | This line is a subclone of HMGUi001-A-8. |
| User feedback | |
Provider |
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| Generator | Institute of Diabetes and Regeneration Research (IDR) |
| Owner | Institute of Diabetes and Regeneration Research (IDR) |
| Distributors | |
| Derivation country | Germany |
External Databases |
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| BioSamples | SAMEA114314345 |
General Information |
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| Publications | |
| * Is the cell line readily obtainable for third parties? |
No |
| Subclone of | |
Donor Information
General Donor Information |
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| Sex | female |
| Ethnicity | Caucasian |
Phenotype and Disease related information (Donor) |
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| Diseases | No disease was diagnosed.
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| Disease associated phenotypes | no phenotypes |
| Family history | no |
| Is the medical history available upon request? | no |
| Is clinical information available? | no |
Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
No
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Other Genotyping (Donor) |
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| Is there genome-wide genotyping or functional data available? |
No
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Donor Relations |
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| Other cell lines of this donor | |
External Databases (Donor) |
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| BioSamples | SAMEA5696688 |
Ethics
Also have a look at the ethics information for the parental line
HMGUi001-A
.
| Is there an MTA available for the cell line? | No |
| For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? | Vectors were generated at IDR, pU6-(BbsI)-sgRNA-CAG-Cas9-Venus-bpA plasmid (Addgene plasmid #86986) was used. |
| Are you aware of any constraints on the use or distribution of the cell line from the owner or any parties identified in the query above? | Yes |
| Constraints for use or distribution | According to donor consent the line must not be used for commercial purposes. For research purposes project-specific review by a medical Ethics Committee is obligatory. |
hIPSC Derivation
General |
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The source cell information can be found in the parental cell line
HMGUi001-A.
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| Passage number reprogrammed | 27 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | HMGUi001-A-8 was targeted with two vectors. For the first vector, the Cas9-Venus-sgRNA vector, a two sgRNAs were cloned into the pU6-(BbsI)-sgRNA-CAG-Cas9-Venus-bpA plasmid (Addgene plasmid #86986). Cas9 and a targeting site specific sgRNAs were expressed from this vector to induce a double strand break at the targeting regions. The second vector contained the template for the intended homology directed repair: the histone 2B (H2B)-Venus-3xHA-tag coding sequence flanked by a the 771 bp upstream of the NEUROD2 start codon and 967 bp downstream of the NEUROD2 stop codon as 5′ and 3′ homology arms (HA). Silent mutations in the sgRNA binding site were introduced to avoid cutting of the correctly inserted sequences. |
Vector free reprogramming |
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Other |
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| Selection criteria for clones | Transfected cells expressed Venus and were sorted by FACS. The genotype of the clones was screened by PCR and verified by Sanger sequencing. |
| Derived under xeno-free conditions |
Yes |
| Derived under GMP? |
No |
| Available as clinical grade? |
No |
Culture Conditions
| Surface coating | Matrigel/Geltrex |
| Feeder cells |
No |
| Passage method | EDTA |
| O2 Concentration | 21 % |
| CO2 Concentration | 5 % |
| Medium |
Other medium:
Base medium: StemMACS iPS-Brew XF medium
Main protein source: xeno-free Serum concentration: 0 % |
| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | Yes |
| Has Rock inhibitor (Y27632) been used at cryo previously with this cell line? | Yes |
| Has Rock inhibitor (Y27632) been used at thaw previously with this cell line? | Yes |
Characterisation
Analysis of Undifferentiated Cells
| Marker | Expressed | Immunostaining | RT-PCR | Flow Cytometry | Enzymatic Assay | Expression Profiles |
| SOX2 |
Yes |
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| POU5F1 (OCT-4) |
Yes |
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| Oct-3 |
Yes |
Differentiation Potency
Microbiology / Virus Screening |
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| Mycoplasma | Negative |
Genotyping
Karyotyping (Cell Line) |
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| Has the cell line karyotype been analysed? |
Yes
46, XX
Karyotyping method:
G-Banding
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Other Genotyping (Cell Line) |
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Genetic Modification
| Genetic modifications not related to a disease |
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