Marciniak Anja, Selck Claudia, Friedrich Betty, Speier Stephan
CRTD - DFG Research Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, Germany ; Paul Langerhans Institute Dresden, German Center for Diabetes Research (DZD), Dresden, Germany.
PLoS One. 2013 Nov 4;8(11):e78706. doi: 10.1371/journal.pone.0078706. eCollection 2013.
Studies on pancreatic cell physiology rely on the investigation of exocrine and endocrine cells in vitro. Particularly, in the case of the exocrine tissue these studies have suffered from a reduced functional viability of acinar cells in culture. As a result not only investigations on dispersed acinar cells and isolated acini were limited in their potential, but also prolonged studies on pancreatic exocrine and endocrine cells in an intact pancreatic tissue environment were unfeasible. To overcome these limitations, we aimed to establish a pancreas tissue slice culture platform to allow long-term studies on exocrine and endocrine cells in the intact pancreatic environment. Mouse pancreas tissue slice morphology was assessed to determine optimal long-term culture settings for intact pancreatic tissue. Utilizing optimized culture conditions, cell specificity and function of exocrine acinar cells and endocrine beta cells were characterized over a culture period of 7 days. We found pancreas tissue slices cultured under optimized conditions to have intact tissue specific morphology for the entire culture period. Amylase positive intact acini were present at all time points of culture and acinar cells displayed a typical strong cell polarity. Amylase release from pancreas tissue slices decreased during culture, but maintained the characteristic bell-shaped dose-response curve to increasing caerulein concentrations and a ca. 4-fold maximal over basal release. Additionally, endocrine beta cell viability and function was well preserved until the end of the observation period. Our results show that the tissue slice culture platform provides unprecedented maintenance of pancreatic tissue specific morphology and function over a culture period for at least 4 days and in part even up to 1 week. This analytical advancement now allows mid -to long-term studies on the cell biology of pancreatic disorder pathogenesis and therapy in an intact surrounding in situ.
胰腺细胞生理学研究依赖于体外对胰腺外分泌细胞和内分泌细胞的研究。特别是对于外分泌组织,这些研究因培养的腺泡细胞功能活力降低而受到影响。结果,不仅对分散的腺泡细胞和分离的腺泡的研究潜力有限,而且在完整胰腺组织环境中对胰腺外分泌和内分泌细胞进行长期研究也不可行。为了克服这些限制,我们旨在建立一个胰腺组织切片培养平台,以便在完整的胰腺环境中对外分泌和内分泌细胞进行长期研究。评估了小鼠胰腺组织切片的形态,以确定完整胰腺组织的最佳长期培养条件。利用优化的培养条件,在7天的培养期内对外分泌腺泡细胞和内分泌β细胞的细胞特异性和功能进行了表征。我们发现,在优化条件下培养的胰腺组织切片在整个培养期内具有完整的组织特异性形态。在培养的所有时间点都存在淀粉酶阳性的完整腺泡,腺泡细胞显示出典型的强细胞极性。胰腺组织切片中的淀粉酶释放量在培养过程中减少,但对增加的蛙皮素浓度保持特征性的钟形剂量反应曲线,且最大释放量比基础释放量高约4倍。此外,直到观察期结束,内分泌β细胞的活力和功能都得到了很好的保留。我们的结果表明,该组织切片培养平台在至少4天甚至部分长达1周的培养期内,能前所未有的维持胰腺组织的特异性形态和功能。这一分析进展现在允许在完整的原位环境中对胰腺疾病发病机制和治疗的细胞生物学进行中长期研究。