Basic study on cryopreservation of rat calvarial osteoblasts with different cryoprotectants.

IF 1.4 4区医学 Q4 CELL BIOLOGY

Cell and Tissue Banking Pub Date : 2024-09-01 Epub Date: 2024-07-08 DOI:10.1007/s10561-024-10142-3

Xu Jiang, Tan Zhijian, Cao Min, Yu Rong, Tan Xinghui, Xin Gong

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Abstract

Cryopreservation is a method adopted for storage of autologous skulls. Herein, this current research sought to explore the effects of different cryoprotectants on the biological characteristics of rat calvarial osteoblasts after cryopreservation. Neonatal Sprague-Dawley rats were selected and their skull tissues were isolated. The skull tissues were allocated into the refrigerating-3M, refrigerating-6M, M199-3M, M199-6M, povidone iodine-3M, and povidone iodine-6M groups according to the usage of cryoprotectants and treatment time (month) and the fresh group. Osteoblasts were isolated from skull tissues in each group through digestion. The histomorphology of the skull was evaluated by H&E staining and cell morphology was observed by microscopy. The viability, proliferation, apoptosis, and osteogenic activity of osteoblasts were assessed by trypan blue staining, MTT, flow cytometry, and alkaline phosphatase (ALP) staining. The skull histomorphology and osteoblast morphology were similar between the fresh and refrigerating groups. Osteoblast viability was weakened after cryopreservation. The longer the refrigeration time, the lower the number of living cells and the higher the apoptosis rate. However, cryopreservation using different cryoprotectants did not evidently affect osteoblast proliferation and ALP activity. Different cryoprotectants show no apparent effect on the osteogenic activity of rat calvarial osteoblasts after cryopreservation.

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使用不同低温保护剂低温保存大鼠钙化成骨细胞的基础研究

低温保存是一种用于储存自体头骨的方法。本研究旨在探讨不同冷冻保护剂对大鼠颅骨骨细胞冷冻保存后生物学特性的影响。研究人员选择了新生的 Sprague-Dawley 大鼠，并分离了它们的头骨组织。根据冷冻保护剂的使用和处理时间（月），将头骨组织分为冷冻-3M、冷冻-6M、M199-3M、M199-6M、聚维酮碘-3M、聚维酮碘-6M 组和新鲜组。各组头骨组织经消化后分离出成骨细胞。头骨的组织形态由 H&E 染色法评估，细胞形态由显微镜观察。通过胰蓝染色、MTT、流式细胞术和碱性磷酸酶（ALP）染色评估成骨细胞的活力、增殖、凋亡和成骨活性。新鲜组和冷藏组的头骨组织形态和成骨细胞形态相似。冷冻保存后，成骨细胞的活力减弱。冷藏时间越长，活细胞数量越少，凋亡率越高。然而，使用不同的冷冻保护剂进行冷冻并没有明显影响成骨细胞的增殖和 ALP 活性。不同的冷冻保护剂对大鼠犊骨成骨细胞冷冻保存后的成骨活性无明显影响。

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来源期刊

Cell and Tissue Banking CELL BIOLOGY-ENGINEERING, BIOMEDICAL

CiteScore

3.10

自引率

13.30%

发文量

审稿时长

6-12 weeks

期刊介绍： Cell and Tissue Banking provides a forum for disseminating information to scientists and clinicians involved in the banking and transplantation of cells and tissues. Cell and Tissue Banking is an international, peer-reviewed journal that publishes original papers in the following areas: basic research concerning general aspects of tissue banking such as quality assurance and control of banked cells/tissues, effects of preservation and sterilisation methods on cells/tissues, biotechnology, etc.; clinical applications of banked cells/tissues; standards of practice in procurement, processing, storage and distribution of cells/tissues; ethical issues; medico-legal issues.