Blood, 1960, Vol. 15, No. 4, pp. 517-524.
© 1960 American Society of Hematology, Inc.
In Vitro and in Vivo Assessment of the Viability
of Dog Marrow after Storage
JOHN A. MANNICK 1,
HARRY L. LOCHTE JR. 1,
E. DONNALL THOMAS 1, and
JOSEPH W. FERREBEE 1
1 Mary Imogene Bassett Hospital (affiliated with Columbia University), Cooperstown, N. Y.
Assessment has been made of the ability of dog marrow cells to survive
storage at 4 C. and at -79 C. Survival has been assessed in vitro by measurements of rates of DNA synthesis and in vivo by ability to restore marrow
functions in lethally irradiated recipients.
Dog bone marrow cells synthesize DNA in vitro at values 30 to 80 per cent of
normal after storage for 24 hours at 4 C. DNA synthesis is reduced to very low
levels by storage for 96 hours at 4 C. Eight lethally irradiated beagle dogs (600
to 1500 r) survived acute irradiation effects and returned to normal health
after receiving intravenous infusions of autologous bone marrow that had been
stored at 4 C. for 24 hours or less. One dog, similarly irradiated, failed to survive when given marrow stored for 96 hours at 4 C. A tenth dog did survive
when given an exceptionally large quantity of marrow stored for 96 hours at
4 C.
Dog marrow cells synthesize DNA in vitro at values 40 to 50 per cent of
normal in samples stored at -79 C. in 15 per cent glycerol for periods up to
17 days. Five lethally irradiated beagle dogs (1000 to 1500 r) survived acute
irradiation effects and returned to normal health after receiving intravenous
infusions of autologous marrow that had been stored at -79 C. in 15 per cent
glycerol. One other dog, similarly irradiated and treated, regained marrow
function but died of hepatitis. Two other irradiated dogs similarly treated did
not regain marrow function and died. These two had received exceptionally
small infusions of marrow cells.
The observations demonstrate that dog marrow can be preserved satisfactorily
at 4 C. for 24 hours and in glycerol at -79 C. for longer periods. The capacity
of the stored marrow to synthesize DNA in vitro has correlated well with its
ability to induce recovery in lethally irradiated autologous recipients. DNA
synthesis in vitro appears to be a useful index of the ability of marrow cells to
proliferate in vivo.
Submitted on July 2, 1959
Accepted on October 15, 1959
