Co-culture of bone marrow mesenchymal stem cells and islet cells influences islet function

doi:10.3969/j.issn.2095-4344.1837

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cells can secrete various growth factors and support hematopoiesis, but whether they can protect and repair the function of islet cells remains unclear.
OBJECTIVE: To investigate the effect of rat bone marrow mesenchymal stem cells co-cultured with islet cells on rat islet function.
METHODS: Bone marrow was extracted from male Sprague-Dawley rats aged 8-9 weeks. Bone marrow mesenchymal stem cells were isolated from bone marrow using density gradient centrifugation combined with adherent culture. Collagenase P (1 g/L, 10 mL) was injected into the common bile duct of another male Sprague-Dawley rats aged 8-9 weeks, and the pancreas was cut off to isolate and purify islet cells using Histopaque discontinuous density gradient. Islet cells with intact cell membrane were selected by hand using 1-mL pipet pipette under microscope, and co-cultured with third-generation bone marrow mesenchymal stem cells. The experiment was divided into control group (single islet cell culture), direct co-culture group (islet cells co-cultured with bone marrow mesenchymal stem cells in culture plates), indirect culture group 1 (indirect co-culture with Transwell dish), and indirect culture group 2 (islet cell culture with bone marrow mesenchymal stem cell medium). The morphology and growth of islets were observed during culture. Insulin secretion was measured by ELISA on the 3rd, 7th, and 14th days after culture. The study protocol was approved by the Ethics Committee of Yanbian University Affiliated Hospital.
RESULTS AND CONCLUSION: The results of variance analysis showed that insulin secretion was significantly increased when islet cells were co-cultured with bone marrow mesenchymal stem cells, and reached a peak on the 7th day of co-culture. Insulin secretion was significantly higher in the indirect co-culture group 1 than the direct co-culture group (P < 0.05). To conclude, the indirect co-culture method of Transwell dish can significantly improve insulin secretion, and the rational co-culture time is 7 days.

Key words: bone marrow mesenchymal stem cells, islet cells, direct co-culture, indirect co-culture, insulin, Sprague-Dawley rats

CLC Number:

Yang Huiming, Yu Xing, Fang Yuhui. Co-culture of bone marrow mesenchymal stem cells and islet cells influences islet function[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(33): 5281-5285.

Figures/Tables 2

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