A comparison of a sucrose-based solution with other preservation media for cold storage of isolated hepatocytes.
A sucrose-based solution has been compared with other preservation solutions (University of Wisconsin (UW) solution and Marshall's citrate solution, with Dulbecco's medium as control) during hypothermic preservation of isolated rat hepatocytes for up to 72 h. Studies on the stability of liver cells at low temperature by exclusion of trypan blue dye and morphological appearance were conducted. During storage beyond 24 h, there was a clear difference between cells stored in Dulbecco's medium and Marshall's citrate and those stored in sucrose-based solution and UW solution, with the former storage groups showing many cells developing large membrane "blebs" and the latter storage groups maintaining a more typical morphology and developing only small membrane protrusions. Dye exclusion was higher in sucrose-based solution (48 h, 75 +/- 7%; 72 h, 65 +/- 6%) and UW solution (48 h, 72 +/- 5%; 72 h, 63 +/- 4%) than in Marshall's citrate (48 h, 31 +/- 5%; 72 h, 10 +/- 1%) and Dulbecco's medium (48 h, 8 +/- 2%; 72 h, 5 +/- 1%). These data suggest that sucrose-based solution should be investigated further as a less complex alternative solution for storage of isolated hepatocytes.
Shanina
I V
IV
Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of the Ukraine, 23 Pereyaslavskaya St., Kharkov 310015, Ukraine.
Kravchenko
L P
LP
Fuller
B J
BJ
Grischenko
V I
VI
D000241:Adenosine; D000493:Allopurinol; D000818:Animals; D003080:Cold Temperature; D004396:Coloring Agents; D005978:Glutathione; D022781:Hepatocytes / Q000166:cytology; D006982:Hypertonic Solutions; D066298:In Vitro Techniques; D007328:Insulin; D019188:Organ Preservation Solutions; D011309:Preservation, Biological / Q000379:methods*; D011887:Raffinose; D051381:Rats; D013395:Sucrose; D014343:Trypan Blue
D004396:Coloring Agents; D006982:Hypertonic Solutions; D007328:Insulin; C054581:Marshall's citrate solution; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D013395:Sucrose; D000493:Allopurinol; D005978:Glutathione; D014343:Trypan Blue; D000241:Adenosine; D011887:Raffinose