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HFF-1
HFF-1
規(guī)格:
貨期:
編號(hào):B167200
品牌:Mingzhoubio

標(biāo)準(zhǔn)菌株
定量菌液
DNA
RNA

規(guī)格:
凍干粉
斜面
甘油
平板 


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      產(chǎn)品名稱
      HFF-1
    
    
    
    
      商品貨號(hào)
      B167200
    
    
      
    
      Organism
      Homo sapiens, human
    
    
      
    
      Tissue
      Skin; foreskin
    
    
      
    
      Cell Type
      Fibroblast
    
    
      
    
      Product Format
      frozen
    
    
      
    
      Morphology
      fibroblast
    
    
      
    
      Culture Properties
      Adherent
    
    
      
    
      Biosafety Level
      1 


    

    Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
    		
    
    
      
    
      Disease
      Normal
    
    
      
    
      Age
      Newborn
    
    
      
    
      Gender
      male
    
    
      
    
      Applications
      
    Can be used to produce feeder cells
    		
    
    
      
    
      Storage Conditions
      Liquid nitrogen vapor phase
    
    
      
    
      Derivation
      
    The cell line was established by ATCC in 2003 from normal human foreskin pooled from two individuals.
    		
    
    
      
    
      Clinical Data
      
    male
    		
    
    
      
    
      Comments
      
    The growth of these cells should be arrested before being used as a feeder layer. ATCC has successfully irradiated (SCRC-1041.1) and treated the cells with Mitomycin C (SCRC-1041.2) for use as a feeder layer. If the HFFs are being used as a feeder layer for ES cells, it is not recommended to use them past passage no. 50 (P50). It is recommended that the feeder cells be plated 24 hours before use at 5X104 cells/cm2 in order to obtain a supportive monolayer for stem cell growth.
    		
    
    
      
    
      Complete Growth Medium
      The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002.  To make the complete growth medium, add the following components to the base medium:  
  • fetal bovine serum to a final concentration of 15%

    • This medium is formulated for use with a 5% CO2 in air atmosphere. (Standard DMEM formulations contain 3.7 g/L sodium bicarbonate and a 10% CO2 in air atmosphere is then recommended).
        
    		
    
    
      
    
      Subculturing
      
    Procedure:
    
    To insure the highest level of viability, be sure to warm media and Trypsin/ EDTA to 37ºC before using it on the cells.
    

    
Cells should be split when they reach confluency. A split ratio of 1:5 to 1:7 is recommended. Volumes used in this protocol are for 225 cm2 (T225); proportionally reduce or increase amount of dissociation medium for culture flasks of other sizes. 
    

      
    
    1. Remove and discard culture medium. 
      2. 
    
    2. Briefly rinse the cell layer with 1X PBS (SCRR-2201) solution to remove all traces of serum, which contain trypsin inhibitor. 
      3. 
    
    3. Add 5 mL of Trypsin-EDTA (0.25% (w/v) Trypsin-0.53 mM EDTA solution, ATCC# 30-2101) solution to flask and incubate for 1 minute, gently tapping the flask observe cells under an inverted microscope until cells detach (usually within 1 to 2 minutes). 
      4. 
    
    4. Add 6.0 to 8.0 mL of complete growth medium and rinse surface of the flask to detach all cells. Gently pipetting up and down will break cell clumps. 
      5. 
    
    5. Transfer all cells into a centrifuge bottle or tube and centrifuge at 270 xg for 5 minutes. 
      6. 
    
    6. Remove and discard the supernatant 
      7. 
    
    7. Add 10 mL complete growth medium to cell pellet and with 10 mL pipette resuspend the cells gently (create a single-cell suspension). 
      8. 
    
    8. Add more complete growth medium to cell suspension as needed to plate cells at approximately 5x106/T225 flask. 
      9. 
    
    9. Place flasks in incubator @ 37°C with a 5% CO2 in air atmosphere.
      10. 

    

    

    
    
        
    
            
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
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      Cryopreservation
      
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    Growth Area (cm2) 
    
            
    
            
    
            
    
            
    
            
    
            
    
            
     
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    		
    
    
      
    
      Culture Conditions
      
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    1xPBS (mL) 
    
            
    
            
    
            
    
            
    
            
    
            
    
            
     
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    		
    
    
      
    
      Name of Depositor
      
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    Trypsin/EDTA (mL) 
    
            
    
            
    
            
    
            
    
            
    
            
    
            
     
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    		
    
    
      
    
      Year of Origin
      
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    Equal vol. Complete Growth Medium (mL)
    
            
    
            
    
            
    
            
    
            
    
            
    
            
     
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    		
    
    
      
    
      References
      
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    Growth Medium (mL) 
    
            
    
            
    
            
    
            
    
            
    
            
    
            
     
    
            
    
            
    
            
    
            
    
            
    
            
    
            
    		
    
    
        

    

						
					
					
    
							
    
							  
							  
							    
    
							      梅經(jīng)理
							      17280875617
							      1438578920
							    
    
							    
    
							      胡經(jīng)理
							      13345964880
							      2438244627
							    
    
							    
    
							      周經(jīng)理
							      17757487661
							      1296385441
							    
    
							    
    
							      于經(jīng)理
							      18067160830
							      2088210172
							    
    
							    
    
							      沈經(jīng)理
							      19548299266
							      2662369050
							    
    
							    
    
							      李經(jīng)理
							      13626845108
							      972239479
							    
    
							    
							      
							
    
					
    
				
				
    
					
				
    
			
			
		
		
    
	
  
    
	






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