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This cell line is a suitable transfection host.

The human diploid fibroblast strain IMR-90 was derived by W.W. Nichols and associates from the lungs of a 16-week female fetus.

16 weeks gestation

Caucasian

female

The division potential, viral susceptibilities and other properties have been thoroughly studied such that the line may be considered as an alternate for WI-38 and other standard human lung cell strains.

The cells have been reported to be capable of attaining 58 population doublings before the onset of senescence.

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning^® T-75 flasks (catalog #430641) are recommended for subculturing this product.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that 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: A subcultivation ratio of 1:2 to 1:8 is recommended

Medium Renewal: Every 3 to 4 days

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

Storage temperature: liquid nitrogen vapor phase

Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

Temperature: 37°C

Amelogenin: X

CSF1PO: 11,13

D13S317: 11,13

D16S539: 10,13

D5S818: 12,13

D7S820: 9,12

THO1: 8,9.3

TPOX: 8,9

vWA: 16,19

G6PD, B