Samsonchi Zakieh, Amirian Roshanak, Tayebi Lobat, Derakhshankhah Hossein, Izadi Zhila, Hajizadeh-Saffar Ensiyeh
Department of Stem Cells and Developmental Biology, Cell Science Research Center, ACECR, Royan Institute for Stem Cell Biology and Technology, Tehran, Iran.
Advanced Therapy Medicinal Product Technology Development Center (ATMP-TDC), Cell Science Research Center, ACECR, Royan Institute for Stem Cell Biology and Technology, Tehran, Iran.
Stem Cell Res Ther. 2025 Mar 13;16(1):136. doi: 10.1186/s13287-025-04168-x.
Diabetes mellitus remains a pervasive global health concern, urging a deeper exploration of islet transplantation as a potential enduring solution. The efficacy of this therapeutic approach pivots on the precision of cryopreservation techniques, ensuring both the viability and accessibility of pancreatic islets. This study delves into the merits of cryopreserving these islets using the disaccharide trehalose, accompanied by an inventive strategy involving poly L proline (PLP) as a cell-penetrating peptide to overcome the cryoprotectant limitations inherent to trehalose.
In our experiments with rat islets, we conducted meticulous viability assessments for fresh and frozen samples. We employed a spectrum of methods, including live/dead staining, insulin/glucagon staining, and measurement of reactive oxygen species (ROS) levels. To gauge functional integrity, we executed glucose-stimulated insulin secretion tests. Subsequently, we transplanted thawed islets into diabetic mice to scrutinize their performance in clinically relevant conditions.
Our study yielded compelling results, affirming the successful cryopreservation of pancreatic islets using trehalose and PLP. Viability, as corroborated through live/dead and insulin/glucagon staining, underscored the sustained preservation of frozen islets. Moreover, these preserved islets exhibited functional integrity by releasing insulin responsively to glucose stimulation. Significantly, upon transplantation into diabetic mice, the thawed islets proficiently restored euglycemia, evidenced by a substantial reduction in fasting blood glucose and an enhanced glucose tolerance.
Our findings accentuate the potential of trehalose and PLP as sophisticated cryoprotectants for preserving pancreatic islets. Beyond highlighting viability and functionality, the preserved islets demonstrated a remarkable capacity to restore euglycemia post-transplantation. This research holds promise in addressing the inherent limitations of islet transplantation, particularly in the realm of Type 1 diabetes treatment.
糖尿病仍然是一个普遍存在的全球健康问题，促使人们更深入地探索胰岛移植作为一种潜在的持久解决方案。这种治疗方法的疗效取决于冷冻保存技术的精确性，以确保胰岛的活力和可用性。本研究探讨了使用二糖海藻糖冷冻保存这些胰岛的优点，并采用了一种创新策略，即将聚L-脯氨酸（PLP）作为细胞穿透肽，以克服海藻糖固有的冷冻保护剂局限性。
在我们对大鼠胰岛的实验中，我们对新鲜和冷冻样本进行了细致的活力评估。我们采用了一系列方法，包括活/死染色、胰岛素/胰高血糖素染色以及活性氧（ROS）水平的测量。为了评估功能完整性，我们进行了葡萄糖刺激胰岛素分泌试验。随后，我们将解冻后的胰岛移植到糖尿病小鼠体内，以在临床相关条件下检查它们的性能。
我们的研究得出了令人信服的结果，证实了使用海藻糖和PLP成功冷冻保存了胰岛。通过活/死和胰岛素/胰高血糖素染色证实的活力强调了冷冻胰岛的持续保存。此外，这些保存的胰岛通过对葡萄糖刺激做出反应释放胰岛素，表现出功能完整性。值得注意的是，移植到糖尿病小鼠体内后，解冻后的胰岛有效地恢复了正常血糖，空腹血糖大幅降低和葡萄糖耐量增强证明了这一点。
我们的研究结果强调了海藻糖和PLP作为保存胰岛的精密冷冻保护剂的潜力。除了突出活力和功能外，保存的胰岛在移植后显示出恢复正常血糖的显著能力。这项研究有望解决胰岛移植的固有局限性，特别是在1型糖尿病治疗领域。
