PittsburghV1, Ncbi, Merge, bibRecord, 000C92

Corneal Stromal Stem Cells versus Corneal Fibroblasts in Generating Structurally Appropriate Corneal Stromal Tissue

Identifieur interne : 000C92 ( Ncbi/Merge ); précédent : 000C91; suivant : 000C93

Corneal Stromal Stem Cells versus Corneal Fibroblasts in Generating Structurally Appropriate Corneal Stromal Tissue

Auteurs : Jian Wu [États-Unis] ; Yiqin Du [États-Unis] ; Mary M. Mann [États-Unis] ; James L. Funderburgh [États-Unis] ; William R. Wagner [États-Unis]

Source :

Experimental eye research [ 0014-4835 ] ; 2014.

RBID : PMC:3979324

Descripteurs français

English descriptors

KwdEn :
- Blotting, Western, Cell Culture Techniques, Corneal Keratocytes (cytology), Corneal Keratocytes (metabolism), Corneal Stroma (cytology), Corneal Stroma (metabolism), Electrophoresis, Polyacrylamide Gel, Extracellular Matrix Proteins, Gene Expression Regulation (physiology), Humans, Microscopy, Electron, Microscopy, Fluorescence, Multiphoton, N-Acetylglucosaminyltransferases (genetics), Phenotype, Proteoglycans (genetics), Real-Time Polymerase Chain Reaction, Stem Cells (cytology), Stem Cells (metabolism), Sulfotransferases (genetics), Tissue Engineering, Tissue Scaffolds.
MESH :
- chemical , genetics : N-Acetylglucosaminyltransferases, Proteoglycans, Sulfotransferases.
- chemical : Extracellular Matrix Proteins.
- cytology : Corneal Keratocytes, Corneal Stroma, Stem Cells.
- metabolism : Corneal Keratocytes, Corneal Stroma, Stem Cells.
- physiology : Gene Expression Regulation.
- Blotting, Western, Cell Culture Techniques, Electrophoresis, Polyacrylamide Gel, Humans, Microscopy, Electron, Microscopy, Fluorescence, Multiphoton, Phenotype, Real-Time Polymerase Chain Reaction, Tissue Engineering, Tissue Scaffolds.

Abstract

Recapitulation of human corneal stromal tissue is believed to be among the most challenging steps in engineering human corneal tissue because of the difficulty in reproducing its highly-ordered hierarchical ultrastructure, which imparts its robust biomechanical properties and optical transparency. In this study, we compared the feasibility of utilizing human corneal stromal stem cells (hCSSCs) and human corneal fibroblasts (hCFs) in the generation of human corneal stromal tissue on a highly-aligned fibrous substrate made from poly(ester urethane) urea. In the serum-free keratocyte differentiation medium supplemented with FGF-2 (10 ng/mL) and TGF-β3 (0.1 ng/mL), hCSSCs successfully differentiated into keratocytes and secreted multilayered lamellae with orthogonally-oriented collagen fibrils, in a pattern mimicking human corneal stromal tissue. The constructs were 60~70 μm thick and abundant in cornea-specific extracellular matrix (ECM) components, including keratan sulfate, lumican, and keratocan. Under the identical conditions, hCFs tended to differentiate into myofibroblasts and deposited a less-organized collagen-fibrillar construct in a pattern with similarities to corneal scar tissue due to a lack of cornea-specific ECM components. These observations demonstrated that hCSSCs showed a much greater potential, under proper substrate and growth factor guidance, to facilitate the generation of a biological human cornea equivalent. Unlike hCSSCs, hCFs were less responsive to these environmental cues and under identical culture conditions generated an ECM that poorly mimicked the native, functional tissue structure and composition.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3979324

DOI: 10.1016/j.exer.2014.01.005
PubMed: 24440595
PubMed Central: 3979324

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Le document en format XML

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<front><div type="abstract" xml:lang="en"><p id="P2">Recapitulation of human corneal stromal tissue is believed to be among the most challenging steps in engineering human corneal tissue because of the difficulty in reproducing its highly-ordered hierarchical ultrastructure, which imparts its robust biomechanical properties and optical transparency. In this study, we compared the feasibility of utilizing human corneal stromal stem cells (hCSSCs) and human corneal fibroblasts (hCFs) in the generation of human corneal stromal tissue on a highly-aligned fibrous substrate made from poly(ester urethane) urea. In the serum-free keratocyte differentiation medium supplemented with FGF-2 (10 ng/mL) and TGF-β3 (0.1 ng/mL), hCSSCs successfully differentiated into keratocytes and secreted multilayered lamellae with orthogonally-oriented collagen fibrils, in a pattern mimicking human corneal stromal tissue. The constructs were 60~70 μm thick and abundant in cornea-specific extracellular matrix (ECM) components, including keratan sulfate, lumican, and keratocan. Under the identical conditions, hCFs tended to differentiate into myofibroblasts and deposited a less-organized collagen-fibrillar construct in a pattern with similarities to corneal scar tissue due to a lack of cornea-specific ECM components. These observations demonstrated that hCSSCs showed a much greater potential, under proper substrate and growth factor guidance, to facilitate the generation of a biological human cornea equivalent. Unlike hCSSCs, hCFs were less responsive to these environmental cues and under identical culture conditions generated an ECM that poorly mimicked the native, functional tissue structure and composition.</p>
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</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A3">Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Mann, Mary M" sort="Mann, Mary M" uniqKey="Mann M" first="Mary M" last="Mann">Mary M. Mann</name>
<affiliation wicri:level="2"><nlm:aff id="A3">Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Funderburgh, James L" sort="Funderburgh, James L" uniqKey="Funderburgh J" first="James L." last="Funderburgh">James L. Funderburgh</name>
<affiliation wicri:level="4"><nlm:aff id="A1">McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A3">Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Wagner, William R" sort="Wagner, William R" uniqKey="Wagner W" first="William R." last="Wagner">William R. Wagner</name>
<affiliation wicri:level="4"><nlm:aff id="A1">McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A2">Department of Surgery, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Surgery, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
<affiliation wicri:level="4"><nlm:aff id="A4">Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">24440595</idno>
<idno type="pmc">3979324</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3979324</idno>
<idno type="RBID">PMC:3979324</idno>
<idno type="doi">10.1016/j.exer.2014.01.005</idno>
<date when="2014">2014</date>
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<idno type="wicri:Area/Pmc/Curation">000E13</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000E13</idno>
<idno type="wicri:Area/Pmc/Checkpoint">001774</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">001774</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Corneal Stromal Stem Cells versus Corneal Fibroblasts in Generating Structurally Appropriate Corneal Stromal Tissue</title>
<author><name sortKey="Wu, Jian" sort="Wu, Jian" uniqKey="Wu J" first="Jian" last="Wu">Jian Wu</name>
<affiliation wicri:level="4"><nlm:aff id="A1">McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A2">Department of Surgery, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Surgery, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Du, Yiqin" sort="Du, Yiqin" uniqKey="Du Y" first="Yiqin" last="Du">Yiqin Du</name>
<affiliation wicri:level="4"><nlm:aff id="A1">McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A3">Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Mann, Mary M" sort="Mann, Mary M" uniqKey="Mann M" first="Mary M" last="Mann">Mary M. Mann</name>
<affiliation wicri:level="2"><nlm:aff id="A3">Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Funderburgh, James L" sort="Funderburgh, James L" uniqKey="Funderburgh J" first="James L." last="Funderburgh">James L. Funderburgh</name>
<affiliation wicri:level="4"><nlm:aff id="A1">McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A3">Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Wagner, William R" sort="Wagner, William R" uniqKey="Wagner W" first="William R." last="Wagner">William R. Wagner</name>
<affiliation wicri:level="4"><nlm:aff id="A1">McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
<affiliation wicri:level="2"><nlm:aff id="A2">Department of Surgery, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Surgery, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
</placeName>
</affiliation>
<affiliation wicri:level="4"><nlm:aff id="A4">Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219, USA</nlm:aff>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
</analytic>
<series><title level="j">Experimental eye research</title>
<idno type="ISSN">0014-4835</idno>
<idno type="eISSN">1096-0007</idno>
<imprint><date when="2014">2014</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p id="P2">Recapitulation of human corneal stromal tissue is believed to be among the most challenging steps in engineering human corneal tissue because of the difficulty in reproducing its highly-ordered hierarchical ultrastructure, which imparts its robust biomechanical properties and optical transparency. In this study, we compared the feasibility of utilizing human corneal stromal stem cells (hCSSCs) and human corneal fibroblasts (hCFs) in the generation of human corneal stromal tissue on a highly-aligned fibrous substrate made from poly(ester urethane) urea. In the serum-free keratocyte differentiation medium supplemented with FGF-2 (10 ng/mL) and TGF-β3 (0.1 ng/mL), hCSSCs successfully differentiated into keratocytes and secreted multilayered lamellae with orthogonally-oriented collagen fibrils, in a pattern mimicking human corneal stromal tissue. The constructs were 60~70 μm thick and abundant in cornea-specific extracellular matrix (ECM) components, including keratan sulfate, lumican, and keratocan. Under the identical conditions, hCFs tended to differentiate into myofibroblasts and deposited a less-organized collagen-fibrillar construct in a pattern with similarities to corneal scar tissue due to a lack of cornea-specific ECM components. These observations demonstrated that hCSSCs showed a much greater potential, under proper substrate and growth factor guidance, to facilitate the generation of a biological human cornea equivalent. Unlike hCSSCs, hCFs were less responsive to these environmental cues and under identical culture conditions generated an ECM that poorly mimicked the native, functional tissue structure and composition.</p>
</div>
</front>
</TEI>
</pmc>
<pubmed><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Corneal stromal stem cells versus corneal fibroblasts in generating structurally appropriate corneal stromal tissue.</title>
<author><name sortKey="Wu, Jian" sort="Wu, Jian" uniqKey="Wu J" first="Jian" last="Wu">Jian Wu</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Du, Yiqin" sort="Du, Yiqin" uniqKey="Du Y" first="Yiqin" last="Du">Yiqin Du</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Mann, Mary M" sort="Mann, Mary M" uniqKey="Mann M" first="Mary M" last="Mann">Mary M. Mann</name>
<affiliation wicri:level="4"><nlm:affiliation>Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Funderburgh, James L" sort="Funderburgh, James L" uniqKey="Funderburgh J" first="James L" last="Funderburgh">James L. Funderburgh</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Wagner, William R" sort="Wagner, William R" uniqKey="Wagner W" first="William R" last="Wagner">William R. Wagner</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA; Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219, USA. Electronic address: wagnerwr@upmc.edu.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA; Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PubMed</idno>
<date when="2014">2014</date>
<idno type="RBID">pubmed:24440595</idno>
<idno type="pmid">24440595</idno>
<idno type="doi">10.1016/j.exer.2014.01.005</idno>
<idno type="wicri:Area/PubMed/Corpus">003686</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">003686</idno>
<idno type="wicri:Area/PubMed/Curation">003663</idno>
<idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">003663</idno>
<idno type="wicri:Area/PubMed/Checkpoint">003663</idno>
<idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">003663</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en">Corneal stromal stem cells versus corneal fibroblasts in generating structurally appropriate corneal stromal tissue.</title>
<author><name sortKey="Wu, Jian" sort="Wu, Jian" uniqKey="Wu J" first="Jian" last="Wu">Jian Wu</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Du, Yiqin" sort="Du, Yiqin" uniqKey="Du Y" first="Yiqin" last="Du">Yiqin Du</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Mann, Mary M" sort="Mann, Mary M" uniqKey="Mann M" first="Mary M" last="Mann">Mary M. Mann</name>
<affiliation wicri:level="4"><nlm:affiliation>Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Funderburgh, James L" sort="Funderburgh, James L" uniqKey="Funderburgh J" first="James L" last="Funderburgh">James L. Funderburgh</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213, USA.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Ophthalmology, University of Pittsburgh, School of Medicine, 203 Lothrop Street, Pittsburgh, PA 15213</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
<author><name sortKey="Wagner, William R" sort="Wagner, William R" uniqKey="Wagner W" first="William R" last="Wagner">William R. Wagner</name>
<affiliation wicri:level="4"><nlm:affiliation>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA; Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219, USA. Electronic address: wagnerwr@upmc.edu.</nlm:affiliation>
<country xml:lang="fr">États-Unis</country>
<wicri:regionArea>McGowan Institute for Regenerative Medicine, University of Pittsburgh, 450 Technology Drive, Suite 300, Pittsburgh, PA 15219, USA; Department of Surgery, University of Pittsburgh, School of Medicine, 200 Lothrop Street, Pittsburgh, PA 15213, USA; Department of Bioengineering, University of Pittsburgh, 300 Technology Drive, Pittsburgh, PA 15219</wicri:regionArea>
<placeName><region type="state">Pennsylvanie</region>
<settlement type="city">Pittsburgh</settlement>
</placeName>
<orgName type="university">Université de Pittsburgh</orgName>
</affiliation>
</author>
</analytic>
<series><title level="j">Experimental eye research</title>
<idno type="eISSN">1096-0007</idno>
<imprint><date when="2014" type="published">2014</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Blotting, Western</term>
<term>Cell Culture Techniques</term>
<term>Corneal Keratocytes (cytology)</term>
<term>Corneal Keratocytes (metabolism)</term>
<term>Corneal Stroma (cytology)</term>
<term>Corneal Stroma (metabolism)</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Extracellular Matrix Proteins</term>
<term>Gene Expression Regulation (physiology)</term>
<term>Humans</term>
<term>Microscopy, Electron</term>
<term>Microscopy, Fluorescence, Multiphoton</term>
<term>N-Acetylglucosaminyltransferases (genetics)</term>
<term>Phenotype</term>
<term>Proteoglycans (genetics)</term>
<term>Real-Time Polymerase Chain Reaction</term>
<term>Stem Cells (cytology)</term>
<term>Stem Cells (metabolism)</term>
<term>Sulfotransferases (genetics)</term>
<term>Tissue Engineering</term>
<term>Tissue Scaffolds</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Cellules souches (cytologie)</term>
<term>Cellules souches (métabolisme)</term>
<term>Humains</term>
<term>Ingénierie tissulaire</term>
<term>Kératocytes cornéens (cytologie)</term>
<term>Kératocytes cornéens (métabolisme)</term>
<term>Microscopie de fluorescence multiphotonique</term>
<term>Microscopie électronique</term>
<term>N-acetylglucosaminyltransferase (génétique)</term>
<term>Phénotype</term>
<term>Protéines de la matrice extracellulaire</term>
<term>Protéoglycanes (génétique)</term>
<term>Réaction de polymérisation en chaine en temps réel</term>
<term>Régulation de l'expression des gènes (physiologie)</term>
<term>Stroma de la cornée (cytologie)</term>
<term>Stroma de la cornée (métabolisme)</term>
<term>Structures d'échafaudage tissulaires</term>
<term>Sulfotransferases (génétique)</term>
<term>Technique de Western</term>
<term>Techniques de culture cellulaire</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>N-Acetylglucosaminyltransferases</term>
<term>Proteoglycans</term>
<term>Sulfotransferases</term>
</keywords>
<keywords scheme="MESH" type="chemical" xml:lang="en"><term>Extracellular Matrix Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="cytologie" xml:lang="fr"><term>Cellules souches</term>
<term>Kératocytes cornéens</term>
<term>Stroma de la cornée</term>
</keywords>
<keywords scheme="MESH" qualifier="cytology" xml:lang="en"><term>Corneal Keratocytes</term>
<term>Corneal Stroma</term>
<term>Stem Cells</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>N-acetylglucosaminyltransferase</term>
<term>Protéoglycanes</term>
<term>Sulfotransferases</term>
</keywords>
<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Corneal Keratocytes</term>
<term>Corneal Stroma</term>
<term>Stem Cells</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Cellules souches</term>
<term>Kératocytes cornéens</term>
<term>Stroma de la cornée</term>
</keywords>
<keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Régulation de l'expression des gènes</term>
</keywords>
<keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Gene Expression Regulation</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Blotting, Western</term>
<term>Cell Culture Techniques</term>
<term>Electrophoresis, Polyacrylamide Gel</term>
<term>Humans</term>
<term>Microscopy, Electron</term>
<term>Microscopy, Fluorescence, Multiphoton</term>
<term>Phenotype</term>
<term>Real-Time Polymerase Chain Reaction</term>
<term>Tissue Engineering</term>
<term>Tissue Scaffolds</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Humains</term>
<term>Ingénierie tissulaire</term>
<term>Microscopie de fluorescence multiphotonique</term>
<term>Microscopie électronique</term>
<term>Phénotype</term>
<term>Protéines de la matrice extracellulaire</term>
<term>Réaction de polymérisation en chaine en temps réel</term>
<term>Structures d'échafaudage tissulaires</term>
<term>Technique de Western</term>
<term>Techniques de culture cellulaire</term>
<term>Électrophorèse sur gel de polyacrylamide</term>
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<front><div type="abstract" xml:lang="en">Recapitulation of human corneal stromal tissue is believed to be among the most challenging steps in engineering human corneal tissue because of the difficulty in reproducing its highly-ordered hierarchical ultrastructure, which imparts its robust biomechanical properties and optical transparency. In this study, we compared the feasibility of utilizing human corneal stromal stem cells (hCSSCs) and human corneal fibroblasts (hCFs) in the generation of human corneal stromal tissue on a highly-aligned fibrous substrate made from poly(ester urethane) urea. In the serum-free keratocyte differentiation medium supplemented with FGF-2 (10 ng/mL) and TGF-β3 (0.1 ng/mL), hCSSCs successfully differentiated into keratocytes and secreted multilayered lamellae with orthogonally-oriented collagen fibrils, in a pattern mimicking human corneal stromal tissue. The constructs were 60-70 μm thick and abundant in cornea-specific extracellular matrix (ECM) components, including keratan sulfate, lumican, and keratocan. Under the identical conditions, hCFs tended to differentiate into myofibroblasts and deposited a less-organized collagen-fibrillar construct in a pattern with similarities to corneal scar tissue due to a lack of cornea-specific ECM components. These observations demonstrated that hCSSCs showed a much greater potential, under proper substrate and growth factor guidance, to facilitate the generation of a biological human cornea equivalent. Unlike hCSSCs, hCFs were less responsive to these environmental cues and under identical culture conditions generated an ECM that poorly mimicked the native, functional tissue structure and composition.</div>
</front>
</TEI>
</pubmed>
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Corneal Stromal Stem Cells versus Corneal Fibroblasts in Generating Structurally Appropriate Corneal Stromal Tissue

Corneal Stromal Stem Cells versus Corneal Fibroblasts in Generating Structurally Appropriate Corneal Stromal Tissue

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