title: Generation of insulin-producing cells from human bone marrow-derived mesenchymal stem cells: comparison of three differentiation protocols.
abstract: BACKGROUND
  Many protocols were utilized for directed differentiation of mesenchymal stem cells (MSCs) to form insulin-producing cells (IPCs). We compared the relative efficiency of three differentiation protocols.
  
  
  METHODS
  Human bone marrow-derived MSCs (HBM-MSCs) were obtained from three insulin-dependent type 2 diabetic patients. Differentiation into IPCs was carried out by three protocols: conophylline-based (one-step protocol), trichostatin-A-based (two-step protocol), and β -mercaptoethanol-based (three-step protocol). At the end of differentiation, cells were evaluated by immunolabeling for insulin production, expression of pancreatic endocrine genes, and release of insulin and c-peptide in response to increasing glucose concentrations.
  
  
  RESULTS
  By immunolabeling, the proportion of generated IPCs was modest ( ≃ 3%) in all the three protocols. All relevant pancreatic endocrine genes, insulin, glucagon, and somatostatin, were expressed. There was a stepwise increase in insulin and c-peptide release in response to glucose challenge, but the released amounts were low when compared with those of pancreatic islets.
  
  
  CONCLUSIONS
  The yield of functional IPCs following directed differentiation of HBM-MSCs was modest and was comparable among the three tested protocols. Protocols for directed differentiation of MSCs need further optimization in order to be clinically meaningful. To this end, addition of an extracellular matrix and/or a suitable template should be attempted.
authors.affiliation: Department of Biotechnology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Mahmoud M
authors.initials: MM
authors.lastname: Gabr
authors.affiliation: Department of Biotechnology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Mahmoud M
authors.initials: MM
authors.lastname: Zakaria
authors.affiliation: Department of Nephrology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Ayman F
authors.initials: AF
authors.lastname: Refaie
authors.affiliation: Department of Pathology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Sherry M
authors.initials: SM
authors.lastname: Khater
authors.affiliation: Department of Pathology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Sylvia A
authors.initials: SA
authors.lastname: Ashamallah
authors.affiliation: Department of Immunology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Amani M
authors.initials: AM
authors.lastname: Ismail
authors.affiliation: Zewail University of Science and Technology, 6th of October City, Giza 12588, Egypt.
authors.forename: Nagwa
authors.initials: N
authors.lastname: El-Badri
authors.affiliation: Department of Urology, Urology and Nephrology Center, Mansoura 35516, Egypt.
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Ghoneim
mesh_terms: D000328:Adult; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002096:C-Peptide / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002478:Cells, Cultured; D005260:Female; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000096:biosynthesis*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008875:Middle Aged; D010641:Phenotype
chemical_list: D002096:C-Peptide; D007328:Insulin; D005947:Glucose