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CHP 3 (M.W.)
CHP 3 (M.W.)
規(guī)格:
貨期:
編號(hào):B164256
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


產(chǎn)品名稱 CHP 3 (M.W.)
商品貨號(hào) B164256
Organism Homo sapiens, human
Tissue skin
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 galactosemia; galactose-1-phosphate uridyl transferase deficient
Age 6 years
Gender male
Ethnicity Black
Storage Conditions liquid nitrogen vapor phase
Karyotype normal human male; stable
Derivation
This fibroblast cell line was established by W.J. Mellman from a skin biopsy of a 6-year-old Black male with galactosemia. 
Comments

This child demonstrated a classic galactosemia picture in infancy and lacks galactose-1-phosphate uridyl transferase activity in his circulating red cells. 

His older sibling (W.W.) lacks the same enzyme but exhibited no clinical symptoms of galactosemia. [Cell line CHP4, ATCC CCL-133]

Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:2 to 1:3
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions

Temperature: 37°C 
Atmosphere: Air, 95%; CO2 5%

Isoenzymes
G6PD, A
Name of Depositor WJ Mellman
Deposited As Homo sapiens
References

. . J. Pediatr. 68: 551, 1966.

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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