LVPEIi006-A

LVIP02-LC12-1

General

Cell Line

hPSCreg name LVPEIi006-A
Cite as:
LVPEIi006-A (RRID:CVCL_D6M4)
Alternative name(s)
LVIP02-LC12-1
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.
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Provider

Generator L.V. Prasad Eye Institute (LVPEI)
Owner L.V. Prasad Eye Institute (LVPEI)
Distributors
Derivation country India

External Databases

BioSamples SAMEA115157827
Cellosaurus CVCL_D6M4

General Information

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

Sex male
Age of donor (at collection) 25-29
Ethnicity Asian, Indian (27 years)

Phenotype and Disease related information (Donor)

Diseases A disease was diagnosed.
The donor/proband is clinically diagnosed with autosomal recessive LCA12 due to a splice disrupting, point mutation in the RD3 gene.
The donor is a carrier of a disease-associated mutation and affected.
Synonyms
  • LCA12
  • Leber congenital amaurosis 12
  • Leber congenital amaurosis caused by mutation in RD3
  • Leber congenital amaurosis type 12
  • RD3 Leber congenital amaurosis
  • amaurosis congenita of Leber, type 12
show more synonyms
Genetic variants
RD3 (target)
1q32.3
NM_001164688.2: c.296+1G>A
NP_001158160.1: p.Arg99*
Homozygous
Disease associated phenotypes
  • RPE pigment migration
  • Sluggish pupil
  • Nystagmus
Is the medical history available upon request? Yes
Is clinical information available? Yes

Karyotyping (Donor)

Has the donor karyotype been analysed?
Yes
Normal Karyotype, 46,XY
Karyotyping method: G-Banding

Other Genotyping (Donor)

Is there genome-wide genotyping or functional data available?
No

Donor Relations

Other cell lines of this donor

External Databases (Donor)

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
Please 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

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.
Source cell origin
An organ that constitutes the external surface of the body. It consists of the epidermis, dermis, and skin appendages.
Synonyms
  • Skin
  • Human Skin
  • SKIN
  • Integument
  • Skin, total
  • Skin, NOS
  • Skin structure (body structure)
  • entire skin
  • skin of body
  • skin organ
show more synonyms
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

Vector type Non-integrating
Vector Episomal
Genes
Is reprogramming vector detectable?
No
Methods used
PCR
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

Type of used vector free reprogramming factor(s)
None

Other

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
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
SOX2
Yes
KLF4
Yes
NANOG
Yes
SSEA-4
Yes
hTERT
Yes
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
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Marker Expressed
GATA6
Yes
AFP
Yes
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Marker Expressed
MSX2
Yes
ACTA2
Yes
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation
Marker Expressed
PAX6
Yes
OTX2
Yes

Microbiology / Virus Screening

Mycoplasma Negative

Genotyping

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
Normal, 46,XY
Passage number: P15
Karyotyping method: G-Banding

Other Genotyping (Cell Line)