LVIP02-LC12-1
LVPEIi006-A
General
Cell Line |
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| hPSCreg name | LVPEIi006-A |
| Cite as: | LVPEIi006-A |
| Alternative name(s) |
LVIP02-LC12-1
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| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Similar lines | No similar lines found. |
| Last update | 12th April 2024 |
| Notes | This is a LCA12 patient-specific, induced pluripotent stem cell line, carrying a homozygous mutation in RD3 gene, at the exon 2 splice donor site. This mutation (c.296+1 G>A) results in splice skipping and inclusion of Intron 2, frameshift, and premature translational termination after codon 99. This iPSC line was generated by the reprogramming of donor skin-derived human dermal fibroblasts (HDFs) using xeno-free reagents and integration-free episomal plasmids viz. pCXLE-hOCT3/4-shp53-F, pCXLE-hSK and pCXLE-hUL (Addgene Plasmids #27077, #27078 and #27080 respectively). This iPSC line maintained proper colony morphology, expressed the stemness and pluripotency markers OCT4, SOX2, NANOG, KLF4, SSEA4, and exhibited a normal karyotype at passage 15. This line efficiently differentiates into all three lineages and could generate retinal organoids and mature retinal pigmented epithelial 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 | SAMEA115157827 |
General Information |
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| Publications | |
| * 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. |
Donor Information
General Donor Information |
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| Sex | male |
| Age of donor (at collection) | 25-29 |
| Ethnicity | Asian, Indian (27 years) |
Phenotype and Disease related information (Donor) |
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| Diseases | A disease was diagnosed.
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| Disease associated phenotypes |
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| Is the medical history available upon request? | Yes |
| Is clinical information available? | Yes |
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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Donor Relations |
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| Other cell lines of this donor | |
External Databases (Donor) |
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| BioSamples | SAMEA115158754 |
Ethics
| Has informed consent been obtained from the donor of the embryo/tissue from which the pluripotent stem cells have been derived? | Yes |
| Was the consent voluntarily given? | Yes |
| Has the donor been informed that participation will not directly influence their personal treatment? | Yes |
| Can you provide us with a copy of the Donor Information Sheet provided to the donor? | No |
| Provide contact information of the holder of the original Donor Information Sheet: | indumathi@lvpei.org |
| Do you (Depositor/Provider) hold the original Donor Consent Form? | Yes |
| Alternatives to consent are available? | No |
| Is there other documentation provided to the donor for consenting purposes? | No |
| Confirm that consent was obtained by a qualified professional | Yes |
| Has the donor agreed to be re-contacted? | Yes |
| Has the donor been informed about how her/his data will be protected? | Yes |
| Please indicate whether the data associated with the donated material has been pseudonymised or anonymised. | pseudonymised |
| Does consent explicitly allow the derivation of pluripotent stem cells? | Yes |
| Does consent expressly prevent the derivation of pluripotent stem cells? | No |
| Does consent pertain to a specific research project? | Yes |
| Details on restriction to research project | Generation of induced pluripotent stem cells(iPSCs) of healthy individuals and patients with inherited eye diseases (IEDs) |
| Does consent permit unforeseen future research, without further consent? | Yes |
| Does the consent permit uses of donated embryo/tissue or derived cell line intended for clinical treatment or human applications? | No |
| Does consent expressly prevent development of commercial products? | No |
| Does consent expressly prevent financial gain from any use of the donated embryo/tissue, including any product made from it? | No |
| Does consent expressly permit storage of donated embryo/tissue for an unlimited time? | Yes |
| Does consent expressly permit storage of cells derived from the donated embryo/tissue for an unlimited time? | Yes |
| Does consent prevent the DONATED BIOSAMPLE from being made available to researchers anywhere in the world? | No |
| Does consent prevent CELLS DERIVED FROM THE DONATED BIOSAMPLE from being made available to researchers anywhere in the world? | No |
Does consent permit research by | |
| an academic institution? | Yes |
| a public organisation? | Yes |
| a non-profit company? | Yes |
| a for-profit corporation? | Yes |
| Does consent expressly permit collection of genetic information? | Yes |
| Does consent expressly permit storage of genetic information? | Yes |
| Does consent prevent dissemination of genetic information? | Yes |
| Has the donor been informed that their donated biosample or derived cells may be tested for the presence of microbiological agents / pathogens? | Yes |
| Has the donor consented to receive information discovered during use of donated embryo/tissue that has significant health implications for the donor? | Yes |
| How may genetic information associated with the cell line be accessed? | Controlled Access |
| Will the donor expect to receive financial benefit, beyond reasonable expenses, in return for donating the biosample? | No |
| Does the consent anticipate that the donor will be notified of results or outcomes of any research involving the donated samples or derived cells? | No |
| Does the consent permit the donor, upon withdrawal of consent, to stop the use of the derived cell line(s) that have already been created from donated samples? | Yes |
| Does the consent permit the donor, upon withdrawal of consent, to stop delivery or use of information and data about the donor? | Yes |
| Does consent permit access to medical records of the donor? | No |
| Does consent permit access to any other source of information about the clinical treatment or health of the donor? | No |
| Has a favourable opinion been obtained from a research ethics committee, or other ethics review panel, in relation to the Research Protocol including the consent provisions? | Yes |
| Name of accrediting authority involved? | IEC, IC-SCR |
| Approval number | LEC-08011; LEC-BHR-P-06-22-878; IC-SCR 04-15-016; 05-21-013 |
| Has a favourable opinion been obtained from a research ethics committee, or other ethics review panel, in relation to the PROPOSED PROJECT, involving use of donated embryo/tissue or derived cells? | Yes |
| Name of accrediting authority involved? | IEC,IC-SCR |
| Approval number | LEC-08011; LEC-BHR-P-06-22-878; IC-SCR 04-15-016; 05-21-013 |
| Do you have obligations to third parties in regard to the use of the cell line? | No |
| Are you aware of any further constraints on the use of the donated embryo/tissue or derived cells? | No |
| 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? | Addgene |
| 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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| Source cell line name | Human dermal fibroblast cells |
| Source cell type |
Dermal fibroblasts are the major cell type in dermis and are commonly accepted as terminally differentiated cells.
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| Source cell origin |
An organ that constitutes the external surface of the body. It consists of the epidermis, dermis, and skin appendages.
Synonyms
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| Source cell type (free text) | Taken from retroauricular surface. |
| Age of donor (at collection) | 25-29 |
| Collected in | 2018 |
| Passage number reprogrammed | P3 |
Reprogramming method |
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| Vector type | Non-integrating |
| Vector | Episomal |
| Genes | |
| Is reprogramming vector detectable? |
No |
| Methods used |
PCR
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| Notes on reprogramming vector detection | Reprogramming vectors were lost at passage 15. Vector backbone specific primer sets spanning the OriP and EBNA1 sequence regions were analyzed by genomic PCR. |
| Files and images showing reprogramming vector expressed or silenced | |
| 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 | Well formed colonies with clear margins and minimal differentiation were manually picked and clonally passaged until passage 5. Stable colonies were further passaged using 1X cell dissociation solution (0.5 mM EDTA and 30 mM NaCl in 1X DPBS) and expanded beyond passage 10 and characterised. |
| Derived under xeno-free conditions |
Yes |
| Derived under GMP? |
No |
| Available as clinical grade? |
Yes |
Culture Conditions
| Surface coating | Vitronectin |
| 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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| KLF4 |
Yes |
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| NANOG |
Yes |
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| SSEA-4 |
Yes |
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| hTERT |
Yes |
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Score:
| Marker | Present | Absent |
| mCpG | ||
| OCT4 |
Morphology pictures
LVPEIi006-A iPSC.tif
Well-formed colonies with clear margins and minimal differentiation were manually picked
Embryoid body formation assay followed by lineage differentiation confirmed the pluripotency of the generated iPSC line.
Differentiation Potency
In vitro spontaneous differentiation
In vitro spontaneous differentiation
Morphology
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
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Other Genotyping (Cell Line) |
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