title: [Protective effect of bone marrow mesenchymal stem cells on islets from hypoxia/reoxygenation-induced injury].
abstract: OBJECTIVE
  To study the protective effects of bone marrow mesenchymal stem cells (BMSCs) of rhesus monkeys on porcine islets from hypoxia/reoxygenation (H/R)-induced injury.
  
  
  METHODS
  BMSCs were isolated and cultured from the marrow of 5 adult rhesus monkeys (weighing, 6-10 kg) by adherent monocytes. Islets were isolated and purified from the pancreas of 5 neonatal porcine (3-5 days old) by collagenase V digestion method, and were cultured with or without BMSCs, and exposed to hypoxia (1% O2) for 12 hours and reoxygenation for 24 or 48 hours, respectively. The experiment was divided into 4 groups: normal islet group (group A), normal islet + BMSCs group (Group B), H/R islet group (group C), and H/R islet + BMSCs group (group D). The survival rate of islets was calculated by fluorescein diacetate/propidium iodide (PI) staining. The viability of the islet cells was detected by cell counting kit 8. Apoptotic rate of islet cells was tested using Annexin V-FITC/PI labeling and flow cytometry. The stimulation index (SI) of islet function was analyzed by glucose-stimulated insulin secretion assay.
  
  
  RESULTS
  The islet cell cluster of group C was more dispersed than that of groups A and B, and group C had more death cells; and the islet cell cluster of group D was more complete and the survival rate was higher than those of group C. The survival rate of islet was 90.2% +/- 9.1%, 88.3% +/- 5.9%, 52.3% +/- 12.1%, and 71.4% +/- 11.5% in groups A, B, C, and D respectively, it was significantly lower in groups C and D than in groups A and B (P < 0.05), but it was significantly higher in group D than in group C (P < 0.05). After coculture of BMSCs and islet at the ratio of 1:10 and 1:20 in group D, the viability of islet cells was significantly higher than that in group C (P < 0.05). The apoptotic rate was 27.1% +/- 3.2%, 24.0% +/- 1.0%, 64.3% +/- 1.8%, and 46.2% +/- 1.4% in groups A, B, C, and D respectively, it was significantly higher in groups C and D than that in groups A and B (P < 0.05), but it was significantly lower in group D than in group C (P < 0.05). There was no significant difference in SI between groups A and B at each time point (P > 0.05), but it was significantly lower in group C than in groups A and B (P < 0.05); and it was significantly higher in group D than in group C at 24 and 72 hours (P < 0.05).
  
  
  CONCLUSIONS
  BMSCs of rhesus monkeys can protect islet vitality and function from H/R-induced injury.
authors.affiliation: Ministry of Health, Regenerative Medicine Research Center, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P R China.
authors.forename: Jiuming
authors.initials: J
authors.lastname: Zhao
authors.forename: Sirong
authors.initials: S
authors.lastname: He
authors.forename: Chengshi
authors.initials: C
authors.lastname: Wang
authors.forename: Jingping
authors.initials: J
authors.lastname: Liu
authors.forename: Shengfu
authors.initials: S
authors.lastname: Li
authors.forename: Younan
authors.initials: Y
authors.lastname: Chen
authors.forename: Yanrong
authors.initials: Y
authors.lastname: Lu
authors.forename: Jingqiu
authors.initials: J
authors.lastname: Cheng
mesh_terms: D000818:Animals; D017209:Apoptosis; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D015687:Cell Hypoxia; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D005260:Female; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D016381:Islets of Langerhans Transplantation; D008253:Macaca mulatta; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000502:physiology*; D010100:Oxygen / Q000008:administration & dosage*; D013552:Swine; D023822:Tissue Engineering / Q000379:methods
chemical_list: D007328:Insulin; D010100:Oxygen