Differentiation of Mesenchymal Stem Cells Derived from Pancreatic Islets and Bone Marrow into Islet-Like Cell Phenotype

BACKGROUND:Regarding regenerative medicine for diabetes, accessible sources of Mesenchymal Stem Cells (MSCs) for induction of insular beta cell differentiation may be as important as mastering the differentiation process itself. METHODOLOGY/PRINCIPAL FINDINGS:In the present work, stem cells from pancreatic islets (human islet-mesenchymal stem cells, HI-MSCs) and from human bone marrow (bone marrow mesenchymal stem cells, BM-MSCs) were cultured in custom-made serum-free medium, using suitable conditions in order to induce differentiation into Islet-like Cells (ILCs). HI-MSCs and BM-MSCs were positive for the MSC markers CD105, CD73, CD90, CD29. Following this induction, HI-MSC and BM-MSC formed evident islet-like structures in the culture flasks. To investigate functional modifications after induction to ILCs, ultrastructural analysis and immunofluorescence were performed. PDX1 (pancreatic duodenal homeobox gene-1), insulin, C peptide and Glut-2 were detected in HI-ILCs whereas BM-ILCs only expressed Glut-2 and insulin. Insulin was also detected in the culture medium following glucose stimulation, confirming an initial differentiation that resulted in glucose-sensitive endocrine secretion. In order to identify proteins that were modified following differentiation from basal MSC (HI-MSCs and BM-MSCs) to their HI-ILCs and BM-ILCs counterparts, proteomic analysis was performed. Three new proteins (APOA1, ATL2 and SODM) were present in both ILC types, while other detected proteins were verified to be unique to the single individual differentiated cells lines. Hierarchical analysis underscored the limited similarities between HI-MSCs and BM-MSCs after induction of differentiation, and the persistence of relevant differences related to cells of different origin. CONCLUSIONS/SIGNIFICANCE:Proteomic analysis highlighted differences in the MSCs according to site of origin, reflecting spontaneous differentiation and commitment. A more detailed understanding of protein assets may provide insights required to master the differentiation process of HI-MSCs to functional beta cells based only upon culture conditioning. These findings may open new strategies for the clinical use of BM-MSCs in diabetes

Isolation and Characterization of Human Trophoblast Side-Population (SP) Cells in Primary Villous Cytotrophoblasts and HTR-8/SVneo Cell Line

Recently, numerous studies have identified that immature cell populations including stem cells and progenitor cells can be found among “side-population” (SP) cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. These features shown by the SP cells suggested that IL7R and IL1R2 are available as markers to detect the SP cells and that vCTB progenitor cells and trophoblast stem cells were involved in the SP cell population

Structure of a novel, macrocyclic Schiff base

(E,E)-1,2,3,4,4a α, 12a β-Hexahydro-7,7,10,- 10-tetramethyl-7H,10H-8,9-dithia-5,12-diazabenzo-cyclodec ene, C14H24N2S2, Mr = 284.49, monoclinic, P2(1)/n, a = 8.962 (1), b = 19.356 (3), c = 10.178 (1) A, β= 114.49 (1) °, V = 1606.7 (7) A3, Z = 4, Dm = 1.21 (1), D chi = 1.176 Mg m−3, μ (Mo K α, λ = 0.71073 A) = 0.31 mm−1, F(000) = 616, T = 296 (1) K, RF = 0.031 for 2066 reflections. The molecule has approximate point symmetry 2 and consists of a 1,2-trans-disubstituted cyclohexane (chair conformation) fused to a ten-membered ring that contains one disulfide and two, trans-substituted, imino groups. Structural parameters associated with the disulfide group, including the C-S-S-C torsion angle [90.8 (1) degrees], are similar to those reported for several acyclic disulfides. The N-C-C-N torsion angle [-63.1 (2) degrees] associated with disubstitution of the cyclohexane ring is substantially larger than those found for protonated or chelated 1,2-trans-diamino-cyclohexane

Structure of 1% copper(II)-doped dichlorobis(1,2-dimethylimidazole)zinc(II)

[Zn<SUB>0.99</SUB>Cu<SUB>0.01</SUB>Cl<SUB>2</SUB>(c<SUB>5</SUB>H<SUB>8</SUB>N<SUB>2</SUB>)<SUB>2</SUB>], M<SUB>r</SUB>= 328.43, monoclinic, P2<SUB>1</SUB>/c, a= 13.857(1), b=7.1324(7), c= 16.356 (2) &#197;, &#946;= 112.87 (1)&#176;, V=1460.4(5) &#197;<SUP>3</SUP>, Z=4, D<SUB>m</SUB> = 1.50(1), D<SUB>x</SUB>= 1.492g cm<SUP>-3</SUP>, &#181;(Mo K&#945;, &#955;= 0.71073 &#197;) = 20.8cm<SUP>-1</SUP>, F(000) = 672.0, T=297(1)K, R=0.028 for 1782 unique observed reflections. The structure contains discrete dichloro-bis(1,2- dimethylimidazole)zinc(II) molecules. Pseudotetrahedral N<SUB>2</SUB>Cl<SUB>2</SUB> coordination about zinc is effected by coordination to two imidazole

Structures of tetrakis(1,2-dimethylimidazole)M<SUP>II</SUP> diperchlorates (M<SUP>II</SUP> = Co, Zn<SUB>0.98</SUB>Co<SUB>0.02</SUB>)

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Structure of triaqua(D,L-β ,-β dimethylcysteato)copper(II) dihydrate, [Cu(C<SUB>5</SUB>H<SUB>9</SUB>NO<SUB>5</SUB>S)(H<SUB>2</SUB>O)<SUB>3</SUB>].2H<SUB>2</SUB>O

This article does not have an abstract