hPSCreg®
This website uses cookies to keep track of login sessions and for internal Matomo Statistics. See our Privacy Policy for details. This website uses cookies, learn more


HMGUi001-A-22

Registration Summary:

NCS1-KO Clone 19

HMGUi001-A-22

General

Cell Line
hPSCreg name HMGUi001-A-22
Cite as:
HMGUi001-A-22
Alternative name(s)
NCS1-KO Clone 19
Cell line type Human induced pluripotent stem cell (hiPSC)
Similar lines
HMGUi001-A
(XM001, BIHi043-A)
HMGUi001-A-8
(C-PEP-mCherry-hiPSC)
HMGUi001-A-43
(hINS-T2A-H2B-Cherry (-/-), hiPSC-INS-T2A-H2B-Cherry reporter, INSCherry/Cherry)
HMGUi001-A-5
(ΔINK4 T2D risk region hiPSC)
HMGUi001-A-42
(NEUROD2 nVenus/nVenus iPSCs)
HMGUi001-A-1
(hINS-T2A-H2B-Cherry (+/-), HMGUi001-A-1)
HMGUi001-A-4
(hiPSC-ARX-T2A-H2B-CFP-Flag)
HMGUi001-A-46
(ARX-CFP/PAX4-mCherry, ARX-T2A-H2B-CFP/H2B-mCherry-RGSHis-T2A-PAX4, ARXnCFP/nCFP/PAX4mCherry/mCherry)
Last update 30th October 2023
User feedback
No feedback available yet.

Login to share your feedback, experiences or results with the research community.

Provider
Generator Max Delbrück Center Berlin Buch (MDC)
Derivation country Germany

External Databases
BioSamples SAMEA114562334

General Information
* Is the cell line readily obtainable for third parties?
Yes
Research use: allowed
Clinical use: not allowed
Commercial use: not allowed
Subclone of

Donor Information

General Donor Information
Sex female
Ethnicity Caucasian

Phenotype and Disease related information (Donor)
Diseases No disease was diagnosed.
Family history no
Is the medical history available upon request? no
Is clinical information available? no

Karyotyping (Donor)
Has the donor karyotype been analysed?
No

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 SAMEA5696688

Ethics

Also have a look at the ethics information for the parental line HMGUi001-A .
Is there an MTA available for the cell line? No
For generation of the cell line, who was the supplier of any recombined DNA vectors or commercial kits used?
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? No

hIPSC Derivation

General
The source cell information can be found in the parental cell line HMGUi001-A.

Reprogramming method
Vector type None

Vector free reprogramming

Other
Derived under xeno-free conditions
Unknown
Derived under GMP?
Unknown
Available as clinical grade?
Unknown

Culture Conditions

Surface coating Matrigel/Geltrex
Feeder cells
No
Passage method Enzyme-free cell dissociation
EDTA
O2 Concentration 5 %
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?
Yes
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
–
NANOG
Yes
–
SSEA-4
Yes
–
TRA 1-60
Yes
–
Score:
Marker Present Absent
mCpG
OCT4
Differentiation Potency
Endoderm
Ont Id: UBERON_0000925
In vitro directed differentiation
Marker Expressed
SOX17
Yes
CXCR4 / CD184 Antigen
Yes
Morphology
20230221KFTF_Trilineage_HMGUi001-A-22_FACS.pdf
FACS results of Trilineage differentiation potential
Mesoderm
Ont Id: UBERON_0000926
In vitro directed differentiation
Marker Expressed
CD140b / PDGFRB (Platelet Derived Growth Factor Receptor Beta)
Yes
CD144 / CDH5 (Cadherin 5)
Yes
Morphology
20230221KFTF_Trilineage_HMGUi001-A-22_FACS.pdf
FACS results Trilineage differentiation potential
Ectoderm
Ont Id: UBERON_0000924
In vitro directed differentiation
Marker Expressed
SOX2
Yes
PAX6
Yes
Morphology
20230221KFTF_Trilineage_HMGUi001-A-22_FACS.pdf
FACS Results Trilineage Differentiation potential

Microbiology / Virus Screening
Mycoplasma Negative

Certificate of Analysis
Is there a certificate of analysis available?
Yes
Passage: 46

Genotyping

Karyotyping (Cell Line)
Has the cell line karyotype been analysed?
Yes
46, XX
Passage number: 49
Karyotyping method: G-Banding

Other Genotyping (Cell Line)

Genetic Modification

Genetic modifications not related to a disease
Neuronal Calcium Sensor 1 (NCS1) (target)
Gene knock-out
9q34.11
The NCS1 gene was genetically engineered using two sgRNAs (5'- CUUGAUGAAGCCUUUGUACC -3' and 5'- UCUGCUGCCUCCAGGUACAA -3') targeting an early, conserved exon 3 with at least 3bp of mismatch to any other site in the human genome to minimize the risk of off-target effects in potential coding regions. Subsequently, 300,000 cells were transfected with ribonucleoprotein (RNP) complexes using the Neon transfection system (Thermo Fisher Scientific) and Neon transfection 10µl kit (Thermo Fisher Scientific). Editing efficiency in the pool was analyzed by Sanger Sequencing and Synthego ICE analysis 48 hours after transfection. Then, single cell clones were generated using automated hiPSC single cell seeding and clonal expansion performed. Clones identified with the desired out-of-frame indel modifications were identified with Sanger Sequencing and ICE analysis.
CRISPR-associated (CRISPR/Cas) System