SYT13-KOnVenus/nVenus-hiPSC
HMGUi001-A-61
General
Cell Line |
|
| hPSCreg name | HMGUi001-A-61 |
| Cite as: | HMGUi001-A-61 |
| Alternative name(s) |
SYT13-KOnVenus/nVenus-hiPSC
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| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines | No similar lines found. |
| Last update | 22nd October 2025 |
| Notes | For the use of the cell line a material transfer agreement with HMGU is obligatory and can be requested from material-transfer@helmholtz-munich.de. |
| 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 | SAMEA119298074 |
General Information |
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| * 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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| Family history | no |
| Is the medical history available upon request? | no |
| Is clinical information available? | no |
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? | Yes |
| For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used? | Vectors were generated at IDR. The 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. An MTA with HMGU is required. |
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 | 26 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | To generate a knock-in/knock-out SYT13KO iPSC line, the complete open reading frame of SYT13 was replaced with an H2B-Venus reporter gene. For the targeting vector, the 3′ (Ex1A, 776 bp) and 5′ (Ex6B, 1070 bp) homology arms were PCR-amplified from genomic DNA purified from HMGUi001-A hiPSCs. These homology arms were inserted into the pKS-H2B-Venus-Intron-pA vector using Gibson assembly. For the Cas9-Venus-sgRNA vector, two sgRNAs targeting exon 1 and exon 6 were designed with the CRISPOR webtool (http://crispor.tefor.net) and cloned into the pU6-(BbsI)-sgRNA-CAG-Cas9-Venus-bpA plasmid (Addgene plasmid #86986) via BbsI digestion followed by Gibson assembly. |
| Genes | |
| Is reprogramming vector detectable? |
No |
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 |
| POU5F1 (OCT-4) |
Yes |
|
Differentiation Potency
In vitro directed differentiation
| Marker | Expressed |
| SOX17 |
Yes |
| FOXA2 |
Yes |
In vitro directed differentiation
| Marker | Expressed |
| SNAIL1 |
Yes |
| SM22-alpha |
Yes |
In vitro directed differentiation
| Marker | Expressed |
| PAX6 |
Yes |
| TUBB3 |
Yes |
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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