A novel BES-gluconate-sucrose (BGS) solution for cold storage of isolated hepatocytes.
We have used hepatocyte suspensions to study how hypothermic storage in a modified University of Wisconsin (mUW) solution affects liver cell metabolism and cell membrane properties. At present the UW solution is the gold standard of organ preservation. However it contains several ingredients which either are expensive or cannot easily be obtained worldwide. The aim of the present study was the development of a novel preservation solution for rat hepatocytes effective and comparable with the mUW, with enhanced buffer capacity and less expensive. In particular, we investigated the effects of the buffering agent BES, a derivate of aminosulfonic acid, on liver cells metabolism during cold storage and rewarming. In the development of this novel preservation solution we have included three key components: gluconate as impermeant anion, sucrose to give additional osmotic support and the aminosulfonic acid BES for its buffer capacity. Our results shown that BGS solution was equally effective to mUW to protect rat hepatocytes against cold preservation injury due to ischemia and reoxigenation. Also, BGS solution is a good alternative with its high buffer capacity, best indices of respiration activity and it is considerably less expensive than the mUW solution. The use of this solution for the storage of isolated hepatocytes may facilitate hepatocyte research in situations in which the more complex and expensive mUW solution is not available since the cost of one liter of BGS is about 1/3 that an equal volume of mUW solution.
Mamprin
Maria Eugenia
ME
Farmacologia and Biologia Molecular, Facultad de Ciencias Bioquimicas y Farmaceuticas, Universidad Nacional de Rosario, Argentina. mmamprin@fbioyf.unr.edu.ar
Petrocelli
Silvana
S
Guibert
Edgardo
E
Rodriguez
Joaquin
J
D017738:Alkanesulfonic Acids; D000818:Animals; D018929:Cell Culture Techniques; D002462:Cell Membrane / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents; D005942:Gluconates; D022781:Hepatocytes / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013395:Sucrose
D017738:Alkanesulfonic Acids; D003451:Cryoprotective Agents; D005942:Gluconates; C012210:BES; D013395:Sucrose; C030691:gluconic acid