C-PEP-mCherry-hiPSC
HMGUi001-A-8
General
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
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| 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 | 23 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | HMGUi001-A was targeted with two vectors. For the first vector, the Cas9-Venus-sgRNA vector, a sgRNA was cloned into the pU6-(BbsI)-sgRNA-CAG-Cas9-Venus-bpA plasmid (Addgene plasmid #86986). Cas9 and a targeting site specific sgRNA were expressed from this vector to induce a double strand break at the targeting region. The second vector contained the template for the intended homology directed repair: the mCherry coding sequence flanked by a 1111 bp left homology arm (5’ HA) and a 901 bp right homology arm (3’ 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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| SSEA-4 |
Yes |
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| TRA 1-60 |
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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