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035 a| (Sirsi) o1001281843;

035 a| (OCoLC)1001281843;

049 a| EREE;

100 1 a| Sellers, Samantha Sue,e| author.?| UNAUTHORIZED;

245 12 a| A quest for articular cartilage progenitor cells leads to temporal regulation of hyaluronan required for 3T3-L1 adipogenesis /c| by Samantha Sue Sellers.;

264 1 a| [Greenville, N.C.] :b| [East Carolina University],c| 2017.;

300 a| 220 pages :b| illustrations (chiefly color).;

336 a| textb| txt2| rdacontent;

337 a| computerb| c2| rdamedia;

338 a| online resourceb| cr2| rdacarrier;

347 a| text fileb| PDFc| 8.407 MB2| rda;

490 1 a| ECU Brody School of Medicine dissertation;

538 a| System requirements: Adobe Reader.;

538 a| Mode of access: World Wide Web.;

502 b| Ph. D.c| East Carolina Universityd| 2017;

500 a| Presented to the faculty of the Department of Anatomy and Cell Biology;

500 a| Advisor: Cheryl B. Knudson;

500 a| Title from PDF t.p. (viewed January 11, 2018).;

520 3 a| During embryogenesis, mesenchymal progenitor cells exhibit hyaluronan-dependent pericellular matrices as a major component of the extracellular matrix. As connective tissues form by cellular condensations, there is a loss of cell-associated hyaluronan to facilitate cell-to-cell adhesion. The goal of this dissertation is to gain insight into how hyaluronan-associated components of the extracellular/pericellular matrix participate in directing differentiation fates of mesenchymal cells. This work began with attempts to isolate and study a primary mesenchymal progenitor population from adult bovine articular cartilage and progressed to the use of the adipocyte-favoring mesenchymal progenitor cell line, 3T3-L1. A particle exclusion assay revealed that 3T3-L1 mesenchymal cells exhibited large hyaluronan-dependent pericellular matrices that were displaced by small hyaluronan fragments consisting of 6-8 disaccharides, suggesting retention of the matrix by a cell surface receptor. These pericellular matrices were enhanced with the addition of exogenous aggrecan (prepared from bovine articular cartilage). However, 3T3-L1 cells lost the ability to synthesize or retain a hyaluronan-dependent pericellular matrix during adipogenesis. We observed a reduction in mRNA and protein expression of CD44, the hyaluronan receptor that anchors the pericellular matrix to the cell surface, during adipogenesis, in addition to a reduction in mRNA expression of Has2 and Vcan. These reductions in pericellular matrix components are likely responsible for the loss of detectable pericellular matrix by 3T3-L1 adipocytes. The importance of CD44's role in pericellular matrix retention was further supported when pericellular matrices could not be formed by 3T3-L1 adipocytes even when exogenous hyaluronan and aggrecan were supplied. Hyaluronan was visualized by staining with a binding protein (HABP) and fluorescent microscopy. Mesenchymal cells exhibited bright cell surface HABP staining whereas adipocytes stained only weakly. However, when the adipocytes were permeabilized with 0.5% Triton-X, hyaluronan was revealed between the lipid droplets of these cells suggesting receptor-mediated endocytosis. When hyaluronan synthesis was blocked with 4-methylumbelliferone, adipogenesis of 3T3-L1 cells was significantly inhibited. Additionally, exogenous aggrecan added to the culture medium during the adipogenic differentiation protocol significantly inhibited adipogenesis. Collectively, these data suggest that 3T3-L1 adipogenesis is dependent upon the temporal regulation of hyaluronan.;

504 a| Includes bibliographical references.;

650 2 a| Hyaluronic Acid.=| ^A1058023;

650 2 a| Adipogenesis.=| ^A1289114;

650 2 a| Cartilage, Articular.=| ^A1037437;

650 2 a| Stem Cells.=| ^A930050;

653 a| 3T3-L1 cells;

653 a| aggrecan;

653 a| extracellular matrix;

700 1 a| Knudson, Cheryl B.,e| degree supervisor.?| UNAUTHORIZED;

710 2 a| East Carolina University.b| Department of Anatomy and Cell Biology.?| UNAUTHORIZED;

830 0 a| ECU Brody School of Medicine dissertation.?| UNAUTHORIZED;

856 40 z| Access via ScholarShipu| http://hdl.handle.net/10342/6366;

949 o| wjh;

994 a| C0b| ERE;

096 a| QU 83;

596 a| 1 4;

998 a| 4733390;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 4733390-1001l| JNETm| JOYNERr| Ys| Yt| JNE3ETDu| 8/16/2017x| ETDz| JERESOURCE;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 4733390-2001l| HSLELECm| HSLr| Ys| Yt| HEETDu| 8/16/2017x| ETDz| HERESOURCE;

A quest for articular cartilage progenitor cells leads to temporal regulation of hyaluronan required for 3T3-L1 adipogenesis / by Samantha Sue Sellers.

Author/creator	Sellers, Samantha Sue author.
Other author	Knudson, Cheryl B., degree supervisor.
Other author	East Carolina University. Department of Anatomy and Cell Biology.
Format	Theses and dissertations
Publication	[Greenville, N.C.] : [East Carolina University], 2017.
Description	220 pages : illustrations (chiefly color).
Supplemental Content	Access via ScholarShip
Subjects	Hyaluronic Acid. Adipogenesis. Cartilage, Articular. Stem Cells.

Series	ECU Brody School of Medicine dissertation ECU Brody School of Medicine dissertation. UNAUTHORIZED
Summary	During embryogenesis, mesenchymal progenitor cells exhibit hyaluronan-dependent pericellular matrices as a major component of the extracellular matrix. As connective tissues form by cellular condensations, there is a loss of cell-associated hyaluronan to facilitate cell-to-cell adhesion. The goal of this dissertation is to gain insight into how hyaluronan-associated components of the extracellular/pericellular matrix participate in directing differentiation fates of mesenchymal cells. This work began with attempts to isolate and study a primary mesenchymal progenitor population from adult bovine articular cartilage and progressed to the use of the adipocyte-favoring mesenchymal progenitor cell line, 3T3-L1. A particle exclusion assay revealed that 3T3-L1 mesenchymal cells exhibited large hyaluronan-dependent pericellular matrices that were displaced by small hyaluronan fragments consisting of 6-8 disaccharides, suggesting retention of the matrix by a cell surface receptor. These pericellular matrices were enhanced with the addition of exogenous aggrecan (prepared from bovine articular cartilage). However, 3T3-L1 cells lost the ability to synthesize or retain a hyaluronan-dependent pericellular matrix during adipogenesis. We observed a reduction in mRNA and protein expression of CD44, the hyaluronan receptor that anchors the pericellular matrix to the cell surface, during adipogenesis, in addition to a reduction in mRNA expression of Has2 and Vcan. These reductions in pericellular matrix components are likely responsible for the loss of detectable pericellular matrix by 3T3-L1 adipocytes. The importance of CD44's role in pericellular matrix retention was further supported when pericellular matrices could not be formed by 3T3-L1 adipocytes even when exogenous hyaluronan and aggrecan were supplied. Hyaluronan was visualized by staining with a binding protein (HABP) and fluorescent microscopy. Mesenchymal cells exhibited bright cell surface HABP staining whereas adipocytes stained only weakly. However, when the adipocytes were permeabilized with 0.5% Triton-X, hyaluronan was revealed between the lipid droplets of these cells suggesting receptor-mediated endocytosis. When hyaluronan synthesis was blocked with 4-methylumbelliferone, adipogenesis of 3T3-L1 cells was significantly inhibited. Additionally, exogenous aggrecan added to the culture medium during the adipogenic differentiation protocol significantly inhibited adipogenesis. Collectively, these data suggest that 3T3-L1 adipogenesis is dependent upon the temporal regulation of hyaluronan.
General note	Presented to the faculty of the Department of Anatomy and Cell Biology
General note	Advisor: Cheryl B. Knudson
General note	Title from PDF t.p. (viewed January 11, 2018).
Dissertation note	Ph. D. East Carolina University 2017
Bibliography note	Includes bibliographical references.
Technical details	System requirements: Adobe Reader.
Technical details	Mode of access: World Wide Web.

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A quest for articular cartilage progenitor cells leads to temporal regulation of hyaluronan required for 3T3-L1 adipogenesis / by Samantha Sue Sellers.

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