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胰岛分离结果受胰腺制备方法的影响。

[Islet isolation outcome is influenced by pancreas preparation method].

作者信息

Pokrywczyńska Marta, Drewa Tomasz, Cieślak Zaneta

机构信息

Uniwersytet Mikołaja Kopernika w Toruniu, Collegium Medicum w Bydgoszczy, Katedra Biologii Medycznej, Zakład Inzynierii Tkankowej.

出版信息

Pol Merkur Lekarski. 2008 Sep;25(147):240-3.

Abstract

UNLABELLED

Pancreatic islet transplantation is a treatment method for type I diabetes. Its outcome is influenced by numerous factors, islet quantity and function being important ones of them.

THE AIM OF THIS STUDY

was to estimate the influence of pancreas preparation method on the outcome of islet isolation in rat.

MATERIAL AND METHODS

6 pancreata harvested from Lewis rats were used in this research. Pancreatic duct was cannulated and pancreas was injected with 1 mg/ml collagenase P solution (Sigma) and then excised. After cutting into smaller fragments, it was digested in collagenase P solution for 15-20 min. Enzyme activity was then stopped by adding dilution medium. Heterogenous cell suspension was centrifuged in density gradient (Gradisol) to isolate islets. Pancreatic islets were collected and islet equivalent was calculated. Islet purity degree was estimated as islet cells to all cells, including exocrine, ratio. Islet viability was estimated using propidium iodide and fluorescein diacetate staining. Photographic documentation was made.

RESULTS

Proper islet morphology, highest number and viability was obtained when pancreas was excised properly (isolation 3 and 4).

CONCLUSION

Pancreas preparation method is one of which influences on islet isolation outcome.

摘要

未标注

胰岛移植是I型糖尿病的一种治疗方法。其结果受多种因素影响,胰岛数量和功能是其中的重要因素。

本研究目的

是评估胰腺制备方法对大鼠胰岛分离结果的影响。

材料与方法

本研究使用从Lewis大鼠获取的6个胰腺。将胰管插管,向胰腺注射1mg/ml的胶原酶P溶液(Sigma公司),然后切除胰腺。切成较小碎片后,在胶原酶P溶液中消化15 - 20分钟。然后通过添加稀释培养基终止酶活性。将异质细胞悬液在密度梯度(Gradisol)中离心以分离胰岛。收集胰岛并计算胰岛当量。胰岛纯度通过胰岛细胞与所有细胞(包括外分泌细胞)的比例来估计。使用碘化丙啶和荧光素二乙酸酯染色评估胰岛活力。进行了摄影记录。

结果

当胰腺切除得当(分离3和4)时,可获得合适的胰岛形态、最高数量和活力。

结论

胰腺制备方法是影响胰岛分离结果的因素之一。

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