LVIP05-RB1-CS1
RUCDRi002-A-63
General
Cell Line |
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| hPSCreg name | RUCDRi002-A-63 |
| Cite as: | RUCDRi002-A-63 (RRID:CVCL_D6N0) |
| Alternative name(s) |
LVIP05-RB1-CS1
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| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines |
RUCDRi002-A (TC-113, 50-001-21, 50-001-21.P20_10, ND50038, LiPSC-GR1.1, TC-1133) WAe001-A-10 (H1_RB1ex3_G3) WAe001-A-11 (H1_RB1ex3_G4) WAe001-A-31 (H1_RB1ex1_D6) WAe009-A-12 (H9_RB1ex3_C7) WAe009-A-13 (H9_RB1ex3_G12LS) RUCDRi002-A-64 (LVIP05-RB1-CS2) RUCDRi002-A-65 (LVIP05-RB1-CS3) WAe001-A-32 (H1_RB1ex1_E9) GENYOi006-A-1 (GRX-MCiPS4F-A2-ETO2-GLIS2) |
| Last update | 13th March 2024 |
| Notes | This is a human induced pluripotent stem cell line generated by CRISPR based editing of a healthy control iPSC line, RUCDRi002-A. It carries compound heterozygous in-del mutations in exon 18 of RB1, leading to frame shift in the reading frame of both the alleles and is predicted to result in premature translation termination and C-terminal truncated protein formation. The line has been stably maintained beyond passage 20, and expresses the stem cell markers OCT4, SOX2, NANOG, KLF4 and SSEA4, and has a normal karyotype. It efficiently differentiates into all germ cell lineages and also forms retinal organoids and mature pigmented RPE cells. |
| User feedback | |
Provider |
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| Generator |
L.V. Prasad Eye Institute (LVPEI)
Contact:
L.V. Prasad Eye Institute (LVPEI) |
| Owner | L.V. Prasad Eye Institute (LVPEI) |
| Distributors | |
| Derivation country | India |
External Databases |
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| BioSamples | SAMEA114430734 |
| Cellosaurus | CVCL_D6N0 |
General Information |
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| Publications |
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| * Is the cell line readily obtainable for third parties? |
Yes Research use: allowed
Clinical use: not allowed
Commercial use: allowed
Additional restrictions:
Terms and conditions apply for commercial use |
| Subclone of | |
Donor Information
General Donor Information |
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| Sex | male |
Phenotype and Disease related information (Donor) |
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| Diseases | No disease was diagnosed.
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| Disease associated phenotypes | no phenotypes |
Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
Yes
Normal karyotype, 46, XY
Karyotyping method:
G-Banding
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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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External Databases (Donor) |
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| BioSamples | SAMEA4938814 |
Ethics
Also have a look at the ethics information for the parental line
RUCDRi002-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? | The gene editing vector plasmid, lentiCRISPRv2, was obtained from Addgene. The RB1 targeting CRISPR guide oligo was then cloned into the BsmBI site, downstream of the U6 promoter. |
| 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 | For research use only |
hIPSC Derivation
General |
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The source cell information can be found in the parental cell line
RUCDRi002-A.
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Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Episomal |
| Is reprogramming vector detectable? |
No |
| Vector map | |
Vector free reprogramming |
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| Type of used vector free reprogramming factor(s) |
None
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Other |
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| Selection criteria for clones | Puromycin Selection |
| 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 |
Enzyme-free cell dissociation
EDTA
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| CO2 Concentration | 5 % |
| Medium | Essential 8™ |
| 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? | No |
| 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 |
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| SOX2 |
Yes |
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| NANOG |
Yes |
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| SSEA-4 |
Yes |
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| cMYC |
Yes |
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| hTERT |
Yes |
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| KLF4 |
Yes |
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Score:
| Marker | Present | Absent |
| mCpG | ||
| OCT4 |
Morphology pictures
Differentiation Potency
In vitro spontaneous differentiation
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
Normal karyotype, 46, XY
Passage number: 15
Karyotyping method:
G-Banding
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Other Genotyping (Cell Line) |
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Genetic Modification
| Disease/phenotype related modifications |
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