CHP 4 (W.W.) (ATCC^® CCL-133^™)

This fibroblast cell line was established by W.J. Mellman from a skin biopsy of an 8-year-old Black male with asymptomatic galactosemia. This child had never exhibited any symptoms of galactosemia although he lacked galactose-1-phosphate uridyl transferase activity in his circulating red cells. Metabolic studies appear to place him in a subgroup of atypical galactosemic patients as described by Segal et al. His younger sibling, (M.W.) demonstrated the classic galactosemic picture in infancy [Cell line CHP3, ATCC CCL-132].

This child had never exhibited any symptoms of galactosemia although he lacked galactose-1-phosphate uridyl transferase activity in his circulating red cells.

This fibroblast cell line was established by W.J. Mellman from a skin biopsy of an 8-year-old Black male with asymptomatic galactosemia.

Metabolic studies appear to place him in a subgroup of atypical galactosemic patients as described by Segal et al.

male

Black

8 years

This fibroblast cell line was established by W.J. Mellman from a skin biopsy of an 8-year-old Black male with asymptomatic galactosemia. This child had never exhibited any symptoms of galactosemia although he lacked galactose-1-phosphate uridyl transferase activity in his circulating red cells. Metabolic studies appear to place him in a subgroup of atypical galactosemic patients as described by Segal et al. His younger sibling, (M.W.) demonstrated the classic galactosemic picture in infancy [Cell line CHP3, ATCC CCL-132].

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended

Protocol: Volumes used in this protocol are for 75 sq cm flasks; 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 Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM 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 37C to facilitate dispersal.
4. Add 2.0 to 3.0 ml of complete growth medium and aspirate cells by gently pipetting
5. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37C.

Freeze medium: complete growth medium, 95%; DMSO, 5%

G6PD, B