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Polyvinyl Pyrrolidone: A Novel Cryoprotectant in Islet Cell Cryopreservation.

Publication ,  Journal Article
El-Shewy, HM; William, FK; Darrabie, M; Collins, BH; Opara, EC
Published in: Cell Transplant
April 2004

The present study was performed on the basis of the hypothesis that the low molecular weight (MW) compounds, DMSO and glycerol, permeate the cell and interact hydrophobically with intracellular proteins, thereby perturbing the cytoskeletal architecture of frozen cells and diminishing islet cell integrity and function. Isolated rat islets were cultured overnight (18-24 h) at 37°C in RPMI medium supplemented with 10% fetal calf serum and 1% mixture of penicillin/streptomycin. Using a programmable temperature controller, samples of precounted islets were then frozen under liquid nitrogen, in the presence of either 2 M DMSO (MW = 0.078 kDa), 3 M glycerol (MW = 0.092 kDa), 5% polyethylene glycol (PEG, MW = 20 kDa), or 10% polyvinylpyrrolidone (PVP, MW = 40 kDa), and stored at -80°C for 1 week. Following thawing and overnight (18-24 h) culture, intact islet recovery was determined by islet counting after dithizone staining. Islet function was assessed by determination of glucose-stimulated insulin secretion in perifusion experiments with Krebs-Ringer bicarbonate buffer, pH 7.4, containing either basal (3.3 mM) or high (16.7 mM) glucose concentrations. The assessment of islet recovery and function of all cryopreserved samples was performed only after thawing and overnight culture (18-24 h) of islets. The mean ± SEM percent intact islet recovery was higher with PVP compared with DMSO (82 ± 4.6 vs. 62.7 ± 3.1%, respectively, p < 0.005, n = 9). Furthermore, the glucose stimulation index of insulin secretion by islets taken from samples frozen with PEG and PVP, after thawing and overnight culture, was comparable to that of freshly isolated islets, in contrast to DMSO and glycerol. There was no significant difference in intact islet recovery and function between samples frozen with PVP and those frozen with PEG. Samples frozen with DMSO and glycerol had similar results in islet recovery and function. These data show that PVP is a new and potent cryoprotectant for islet cell freezing.

Duke Scholars

Published In

Cell Transplant

DOI

EISSN

1555-3892

Publication Date

April 2004

Volume

13

Issue

3

Start / End Page

237 / 243

Location

United States

Related Subject Headings

  • Time Factors
  • Temperature
  • Rats, Sprague-Dawley
  • Rats
  • Povidone
  • Pharmaceutic Aids
  • Nitrogen
  • Neurology & Neurosurgery
  • Male
  • Islets of Langerhans
 

Citation

APA
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ICMJE
MLA
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El-Shewy, H. M., William, F. K., Darrabie, M., Collins, B. H., & Opara, E. C. (2004). Polyvinyl Pyrrolidone: A Novel Cryoprotectant in Islet Cell Cryopreservation. Cell Transplant, 13(3), 237–243. https://doi.org/10.3727/000000004783983927
El-Shewy, Hesham M., F Kendall William, Marcus Darrabie, Bradley H. Collins, and Emmanuel C. Opara. “Polyvinyl Pyrrolidone: A Novel Cryoprotectant in Islet Cell Cryopreservation.Cell Transplant 13, no. 3 (April 2004): 237–43. https://doi.org/10.3727/000000004783983927.
El-Shewy HM, William FK, Darrabie M, Collins BH, Opara EC. Polyvinyl Pyrrolidone: A Novel Cryoprotectant in Islet Cell Cryopreservation. Cell Transplant. 2004 Apr;13(3):237–43.
El-Shewy, Hesham M., et al. “Polyvinyl Pyrrolidone: A Novel Cryoprotectant in Islet Cell Cryopreservation.Cell Transplant, vol. 13, no. 3, Apr. 2004, pp. 237–43. Pubmed, doi:10.3727/000000004783983927.
El-Shewy HM, William FK, Darrabie M, Collins BH, Opara EC. Polyvinyl Pyrrolidone: A Novel Cryoprotectant in Islet Cell Cryopreservation. Cell Transplant. 2004 Apr;13(3):237–243.

Published In

Cell Transplant

DOI

EISSN

1555-3892

Publication Date

April 2004

Volume

13

Issue

3

Start / End Page

237 / 243

Location

United States

Related Subject Headings

  • Time Factors
  • Temperature
  • Rats, Sprague-Dawley
  • Rats
  • Povidone
  • Pharmaceutic Aids
  • Nitrogen
  • Neurology & Neurosurgery
  • Male
  • Islets of Langerhans