Improved long-term storage of hybridomas at -80 degrees C using a bovine milk derivative.
A medium comprising 40% bovine milk fraction and 10% DMSO (medium A) was used for the long-term storage of hybridomas at -80 degrees C. The viability of the cells, their growth recovery and ability to secrete antibody were studied and the results were compared to those obtained after storage in a medium containing 40% fetal calf serum and 10% of DMSO (medium B). Hybridomas have been kept for 2 years in medium A; the viability of such cells was 75%, the cells were healthy (electron microscopy), they rapidly proliferated when they were cultured in RPMI supplemented with 10% FCS or with 9% milk fraction + 1% FCS and they released measurable levels of antibody. In contrast, hybridomas stored under the same conditions but in medium B died after 6 months.
Derouiche
F
F
Biochemistry Laboratory School of Medicine, Faculty of Sciences, Nancy University I, Vandoeuvre, France.
Bour
J M
JM
Legrand
C
C
Capiaumont
J
J
Belleville
F
F
Linden
G
G
Nabet
P
P
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002455:Cell Division; D002470:Cell Survival; D003451:Cryoprotective Agents; D005615:Freezing; D006825:Hybridomas; D066298:In Vitro Techniques; D051379:Mice; D008854:Microscopy, Electron; D008892:Milk; D008970:Molecular Weight; D011309:Preservation, Biological / Q000379:methods*; D013997:Time Factors
D000911:Antibodies, Monoclonal; D003451:Cryoprotective Agents