hiPSC-F2-3F1
LVPEIi001-A
General#
Cell Line |
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| hPSCreg Name | LVPEIi001-A |
| Alternative name(s) |
hiPSC-F2-3F1
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| Cell line type | Human induced pluripotent stem cell (hiPSC) |
| Last update | 23rd April 2022 |
| Notes | This is a normal control human iPSC line generated from the dermal fibroblast of an healthy volunteer using retroviral vectors encoding hOCT3/4, hSOX2, hKLF4 and hcMYC transgenes. This line has been stably maintained beyond passage 20, expresses the endogenous stem cell markers such as, OCT4, SOX2, NANOG, SSEA4 and ALP, exhibits a normal karyotype, forms teratomas comprising of all three lineage cells and can efficiently differentiate into brain and eye lineage. It can efficiently generate retinal pigmented epithelium and corneal epithelium as uniform monolayer sheets in 2D cultures and can form brain, retinal and corneal organoids in 3D cultures. |
| User feedback | |
Provider |
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| Generator |
LV Prasad Eye Institute (LVPEI)
Contact:
LV Prasad Eye Institute (LVPEI) |
| Owner | LV Prasad Eye Institute (LVPEI) |
| Distributors | |
| Derivation country | India |
External Databases |
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| BioSamples | SAMEA14207085 |
General Information |
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| Publications |
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| * Is the cell line readily obtainable for third parties? |
Yes Research: allowed
Clinical: not allowed
Commercial: not allowed
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Donor Information#
General Donor Information |
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| Sex | female |
| Age of donor (at collection) | 35-39 |
| Ethnicity | Indian (35 years old) |
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 inherited or systemic disease pathology reported. |
Karyotyping (Donor) |
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| Has the donor karyotype been analysed? |
Yes
Normal Karyotype, 46, XX
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 | SAMEA14207086 |
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 | Generating GMP grade, human induced pluripotent stem cell (iPSC) lines and retinal cells for pre-clinical and clinical evaluations. |
| 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? | Yes |
| 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? | Yes |
| 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, IBSC |
| Approval number | LEC-08011, IC-SCR-05-21-013, IBSC-06-21-009 |
| 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, IBSC |
| Approval number | LEC-08011, IC-SCR-05-21-013, IBSC-06-21-009 |
| 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 type |
dermal fibroblastDermal fibroblasts are the major cell type in dermis and are commonly accepted as terminally differentiated cells.
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| Source cell origin |
SkinAn 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) | 35-39 |
| Collected in | 2011 |
| Passage number reprogrammed | P3 |
Reprogramming method |
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| Vector type | Integrating |
| Vector | Virus (Retrovirus) |
| Genes | |
| Is the used vector excisable? |
No |
| Absence of reprogramming vector(s)? |
No |
| Reprogramming vectors silenced? |
Unknown |
| Methods used |
Immunostaining, PCR, RT-PCR
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| Notes on reprogramming vector silencing | Myc transgene integration and expression is not detected |
| Files and images showing reprogramming vector expressed or silenced | |
| Vector map | |
Vector free reprogramming |
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Other |
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| Selection criteria for clones | Manual picking and passaging till Passage 5. The stably expanding clones are subsequently passaged as cell suspensions prepared using 1X cell dissociation solution (1X DPBS containing 0.5 mM EDTA and 30 mM NaCl) treatment for 6-7 minutes. |
| Derived under xeno-free conditions |
No |
| 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 | mTeSR™ 1 |
| Has Rock inhibitor (Y27632) been used at passage previously with this cell line? | No |
| 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 | FACS | 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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| Alkaline Phosphatase |
Yes |
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Teratoma assay confirmed three lineage differentiation and pluripotency of the iPSC line.
Differentiation Potency
Microbiology / Virus Screening |
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| HIV 1 | Negative |
| HIV 2 | Negative |
| Hepatitis B | Negative |
| Hepatitis C | Negative |
| Mycoplasma | Negative |
Genotyping#
Karyotyping (Cell Line) |
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| Has the cell line karyotype been analysed? |
Yes
46, XX
Passage number: 20
Karyotyping method:
G-Banding
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Other Genotyping (Cell Line) |
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