RUCDRi002-A-63

LVIP05-RB1-CS1

General

Cell Line

hPSCreg name RUCDRi002-A-63
Cite as:
RUCDRi002-A-63
Alternative name(s)
LVIP05-RB1-CS1
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.
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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 SAMEA114430734

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

Subclone of

Donor Information

General Donor Information

Sex male

Phenotype and Disease related information (Donor)

Diseases No disease was diagnosed.
Disease associated phenotypes no phenotypes

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

External Databases (Donor)

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

The source cell information can be found in the parental cell line RUCDRi002-A.

Reprogramming method

Vector type Non-integrating
Vector Episomal
Is reprogramming vector detectable?
No
Vector map

Vector free reprogramming

Type of used vector free reprogramming factor(s)
None

Other

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
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
NANOG
Yes
SSEA-4
Yes
cMYC
Yes
hTERT
Yes
KLF4
Yes
Score:
Marker Present Absent
mCpG
OCT4
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro spontaneous differentiation
Marker Expressed
GATA6
Yes
FOXA1
Yes
Mesoderm
Ont Id: UBERON_0000926
In vitro spontaneous differentiation
Marker Expressed
ACTA2
Yes
MSX2
Yes
Ectoderm
Ont Id: UBERON_0000924
In vitro spontaneous differentiation
Marker Expressed
PAX6
Yes
hMAP2
Yes

Microbiology / Virus Screening

Mycoplasma Negative

Genotyping

Karyotyping (Cell Line)

Has the cell line karyotype been analysed?
Yes
Normal karyotype, 46, XY
Passage number: 15
Karyotyping method: G-Banding

Other Genotyping (Cell Line)

Genetic Modification

Disease/phenotype related modifications
Targeted in-del mutations within Exon 18 of RB1. The lentiCRISPRv2 construct was obtained from Addgene. The 20 mer CRISPR guide target region and PAM sequence within Exon 18 of RB1 is: 5'-AAACAATCAAAGGACCGAGA-AGG-3’. The DNA guide oligos for both strands were designed, anealed and cloned at the BsmBI site, downstream of the U6 promoter of lentiCRISPRv2. The guide construct was nucleofected into NcGMP1 cells and the edited cells were clonally expanded, screened by targeted sequencing. Clones with the desired in-del edits that disrupt RB1 protein expression were expanded and characterized.
Synonyms
  • RB
  • retinoblastoma
  • RETINOBLASTOMA, MALIGNANT
  • Retinoblastoma
  • Retinoblastoma, NOS
show more synonyms
Genetic modifications
RB1 (target)
Gene knock-out
chr13:48453017
The edits in RB1 gene are as follows: Allele 1: c. [1736_1745del10] Allele 2: c.[1736_1737insA]
CRISPR-associated (CRISPR/Cas) System