title: Characterization of Insulin-Secreting Porcine Bone Marrow Stromal Cells Ex Vivo and Autologous Cell Therapy In Vivo.
abstract: Cell therapy could potentially meet the need for pancreas and islet transplantations in diabetes mellitus that far exceeds the number of available donors. Bone marrow stromal cells are widely used in clinical trials mainly for their immunomodulatory effects with a record of safety. However, less focus has been paid to developing these cells for insulin secretion by transfection. Although murine models of diabetes have been extensively used in gene and cell therapy research, few studies have shown efficacy in large preclinical animal models. Here we report optimized conditions for ex vivo expansion and characterization of porcine bone marrow stromal cells and their permissive expression of a transfected insulin gene. Our data show that these cells resemble human bone marrow stromal cells in surface antigen expression, are homogeneous, and can be reproducibly isolated from outbred Yorkshire-Landrace pigs. Porcine bone marrow stromal cells were efficiently expanded in vitro to >10(10) cells from 20 ml of bone marrow and remained karyotypically normal during expansion. These cells were electroporated with an insulin expression plasmid vector with high efficiency and viability, and secreted human insulin and C-peptide indicating appropriate processing of proinsulin. We showed that autologous insulin-secreting bone marrow stromal cells implanted and engrafted in the liver of a streptozotocin-diabetic pig that modeled type 1 diabetes resulted in partial, but significant, improvement in hyperglycemia that could not be ascribed to regeneration of endogenous β-cells. Glucose-stimulated insulin secretion in vivo from implanted cells in the treated pig was documented by a rise in serum human C-peptide levels during intravenous glucose tolerance tests. Compared to a sham-treated control pig, this resulted in significantly reduced fasting hyperglycemia, a slower rise in serum fructosamine, and prevented weight loss. Taken together, this study suggests that bone marrow stromal cells merit further development as autologous cell therapy for diabetes.
authors.affiliation: Division of Medical Sciences, Humphrey Oei Institute of Cancer Research, National Cancer Centre, Singapore, Republic of Singapore.
authors.forename: Hai Van Thi
authors.initials: HV
authors.lastname: Do
authors.forename: Wan Ting
authors.initials: WT
authors.lastname: Loke
authors.forename: Irene
authors.initials: I
authors.lastname: Kee
authors.forename: Vivienne
authors.initials: V
authors.lastname: Liang
authors.forename: Sebastian J
authors.initials: SJ
authors.lastname: David
authors.forename: Shu Uin
authors.initials: SU
authors.lastname: Gan
authors.forename: Sze Sing
authors.initials: SS
authors.lastname: Lee
authors.forename: Wai Har
authors.initials: WH
authors.lastname: Ng
authors.forename: Heng Nung
authors.initials: HN
authors.lastname: Koong
authors.forename: Hock Soo
authors.initials: HS
authors.lastname: Ong
authors.forename: Kok Onn
authors.initials: KO
authors.lastname: Lee
authors.forename: Roy Y
authors.initials: RY
authors.lastname: Calne
authors.forename: Oi Lian
authors.initials: OL
authors.lastname: Kon
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D064987:Cell- and Tissue-Based Therapy; D003921:Diabetes Mellitus, Experimental; D004195:Disease Models, Animal; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D013552:Swine
chemical_list: D007328:Insulin