LVIP05-RB1-CS3	
    			    			
                RUCDRi002-A-65            
            
        General
Cell Line | 
					|
| hPSCreg name | RUCDRi002-A-65 | 
| Cite as: | RUCDRi002-A-65 (RRID:CVCL_D6N2) | 
| Alternative name(s) | 
									 
	LVIP05-RB1-CS3	 
							 | 
						
| 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) RUCDRi002-A-63 (LVIP05-RB1-CS1) RUCDRi002-A-64 (LVIP05-RB1-CS2) WAe001-A-10 (H1_RB1ex3_G3) WAe001-A-11 (H1_RB1ex3_G4) WAe001-A-31 (H1_RB1ex1_D6) 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 homozygous in-del mutation in exon 18 of RB1, leading to frame shift in the reading frame 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 can efficiently differentiate into all germ cell lineages. | 
| User feedback | |
Provider | 
					|
| 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 | SAMEA115170104 | 
| Cellosaurus | CVCL_D6N2 | 
							    
							General Information | 
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| Publications | 
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| * 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 | 
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| Sex | male | 
Phenotype and Disease related information (Donor) | 
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| Diseases | No disease was diagnosed.
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Karyotyping (Donor) | 
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| Has the donor karyotype been analysed? | 
								 Yes 								
																	
												Normal karyotype, 46, XY
											 
																				
																															
											Karyotyping method:
											G-Banding											
																						 
																			 | 
						
Other Genotyping (Donor) | 
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| 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 | 
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| 
						 The source cell information can be found in the parental cell line
						RUCDRi002-A. 
					 | 
				|
| Passage number reprogrammed | 35 | 
Reprogramming method | 
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| Vector type | Non-integrating | 
| Vector | Episomal | 
| 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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| 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 						                 | 
																                
						                
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| SOX2 | 
											 Yes 						                 | 
																                
						                
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| NANOG | 
											 Yes 						                 | 
																                
						                
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| KLF4 | 
											 Yes 						                 | 
																                
						                
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| cMYC | 
											 Yes 						                 | 
																                
						                
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| SSEA-4 | 
											 Yes 						                 | 
																                
						                
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| hTERT | 
											 Yes 						                 | 
																                
						                
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Morphology pictures
Differentiation Potency
In vitro spontaneous differentiation
					In vitro spontaneous differentiation
					Morphology
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) | 
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Genetic Modification
| Disease/phenotype related modifications | 
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