A simple method for cryopreservation of purified adult pig pancreatic cells.
Cryopreservation of islet cells may benefit many aspects of islet transplantation including storage, transport of islets, immunomodulation, reduction of the exocrine contaminants. Isolation and purification of islets from large mammalian pancreases have been encountered by many problems. We reported an autodigestion method for preparation of adult pig pancreatic cells. In this paper, we describe a newly invented simple method for cryopreservation of purified adult pig pancreatic cells. The viability and function of islet cells before (precryo.) and after (postcryo.) the cryopreservation procedure were compared. Islet cell harvested by the autodigestion method were suspended in cryoprotectant "Cell banker." A biofreezing vessel "Bicell" containing a cryogenic vial with cell suspension was frozen at -80 degrees C. After 4 wk, the cryogenic vial was rapidly thawed at 37 degrees C water, the islet cells were washed and cultured for overnight. Number of dithizone-stained precryo. and postcryo. cells were 1.71 +/- 0.5 x 10(5) and 1.75 +/- 0.98 x 10(5) cells/pancreas respectively; therefore, the viability was 99%. Insulin release following high and low concentrations of glucose in precryo. (12.02 +/- 3.63 ng/ml: low-glucose, 9.81 +/- 3.59 ng/ml: high glucose) and precryo. (5.57 +/- 4.03 ng/ml: low-glucose, 10.24 +/- 8.88 ng/ml: high glucose) cells showed a marked improvement in response in postcryo. cells. The insulin content of precryo. and postcryo. cells were 914.4 +/- 277.4 and 93.6 +/- 15.4 ng/ml, respectively. This simple and efficient cryopreservation protocol may be applied for a large-scale storage of adult pig islet cells to establish an islet bank for transplantation.
Department of Surgery III, Tokyo Women's Medical College, Japan.
S
S
Sato
H
H
Ohgawara
Y
Y
Nakagawa
N
N
Katagiri
S
S
Fuchinoue
S
S
Teraoka
E
E
Aikawa
K
K
Ota
D000367:Age Factors; D000818:Animals; D002469:Cell Separation; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D004230:Dithizone; D016381:Islets of Langerhans Transplantation / Q000379:methods*; D013552:Swine; D014015:Tissue Banks
D004230:Dithizone