J Korean Soc Endocrinol.  2006 Feb;21(1):22-31. 10.3803/jkes.2006.21.1.22.

Standardization of Isolation Procedure and Analysis of Variables on Successful Isolation of Islet from the Human Pancreas

• Kim SC
• Han DJ
• Kim IH
• We YM
• Kim YH
• Kim JH
• Back JH
• Lim DG

• KMID: 2200654
• DOI: http://doi.org/10.3803/jkes.2006.21.1.22

Abstract

BACKGROUND: Identifying the donor and isolation-related factors during the islet isolation would be greatly helpful to improve the result of human islet isolation for successful clinical islet transplantation.
METHODS
Sixty-nine pancreata from cadaveric donors were isolated with standard protocol and analyzed to identify the donor factors and isolation variables for successful isolation. Islet isolations recovered > or = 100,000 Islet Equivalent (IEQ, n=53) were compared to islet mass less than 100,000 IEQ (n=16).
RESULTS
The mean islet recovery was 216.0 x 10(3) +/- 173.7 x 10(3) (IEQ) before purification and 130.6 x 10(3) +/- 140.2 x 10(3) (IEQ) after purification. Mean purity was 54 +/- 31%. Mean age of donor was 31.2 +/- 13.2 year and mean cold ischemic time was 6.9 +/- 6.2 hour. Quality of isolated islets was acceptable in terms of bacterial culture, viability and secretory function in vitro and in vivo. In univariate analysis on successful isolation, status of pancreas was the only significant factor and sex, duration of collagenase expansion and digestion time were marginal factors. Stepwise multivariate logistic regression analysis showed donor sex, status of pancreas and digestion time were significant factors for the successful islet isolation.
CONCLUSION
This study confirms some donor factors and variables in isolation process can influence the ability to obtain the successful isolation of human islet. Enough experiences and pertinent review of donor and isolation factors can make islet isolation successful, supporting the clinical islet transplantation without spending of cost.

MeSH Terms

Cadaver
Cold Ischemia
Collagenases
Digestion
Humans*
Islets of Langerhans Transplantation
Logistic Models
Pancreas*
Tissue Donors
Collagenases

Figure

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  Fig. 1 Schematic diagram on islet isolation and transplantation of human pancreas islet cell.

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  Fig. 2 Chronological change of islet yield (IEQ) from human cadaveric pancreas. It shows gradual increase of islet yield since first year of isolation. Shaded bar shows islet yield after purification.

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  Fig. 3 (A) Glucose control after islet transplantation into nude mice in representative cases. Blood glucose level was elevated to more than 400 mg/dL after nephrectomy of islet-bearing kidney. (B) Cure rate of diabetes in nude mice according to number of transplanted islets. Euglycemia was obtained in 30% of nude mice for 750 (IEQ), 40% for 1,000 (IEQ), and 90% for 2,000 (IEQ) transplantation.

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