Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells
Mesenchymal stromal cells (MSC) are differentiation competent cells and may generate, among others, mature osteoblasts or chondrocytes<em> in vitro</em> and <em>in vivo</em>. Laminin-5 and type I collagen are important components of the extracellular matrix. They are involved...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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Open Medical Publishing
2012-12-01
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| Series: | Orthopedic Reviews |
| Subjects: | |
| Online Access: | http://www.pagepress.org/journals/index.php/or/article/view/4467 |
| _version_ | 1818878512611721216 |
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| author | Falk Mittag Eva-Maria Falkenberg Alexandra Janczyk Marco Götze Tino Felka Wilhelm K. Aicher Torsten Kluba |
| author_facet | Falk Mittag Eva-Maria Falkenberg Alexandra Janczyk Marco Götze Tino Felka Wilhelm K. Aicher Torsten Kluba |
| author_sort | Falk Mittag |
| collection | DOAJ |
| description | Mesenchymal stromal cells (MSC) are differentiation competent cells and may generate, among others, mature osteoblasts or chondrocytes<em> in vitro</em> and <em>in vivo</em>. Laminin-5 and type I collagen are important components of the extracellular matrix. They are involved in a variety of cellular and extracellular activities including cell attachment and osteogenic differentiation of MSC. MSC were isolated and expanded using media conforming good medical practice (GMP)-regulations for medical products. Cells were characterized according to the defined minimal criteria for multipotent MSC. MTT- and BrdU-assays were performed to evaluate protein-dependent (laminin-5, laminin-1, type I collagen) metabolic activity and proliferation of MSC. MSC-attachment assays were performed using protein-coated culture plates. Osteogenic differentiation of MSC was measured by protein-dependant mineralization and expression of osteogenic marker genes (osteopontin, alkaline phophatase, Runx2) after three, seven and 28 days of differentiation. Marker genes were identified using quantitative reverse-transcription polymerase chain reaction. Expansion of MSC in GMP-conforming media yielded vital cells meeting all minimal criteria for MSC. Attachment assay revealed a favorable binding of MSC to laminin-5 and type I collagen at a protein concentration of 1-5 fmol/mL. Compared to plastic, osteogenic differentiation was significantly increased by laminin-5 after 28 days of culture (P<0.04). No significant differences in gene expression patterns were observed. We conclude that laminin-5 and type I collagen promote attachment, but laminin-1 and laminin-5 promote osteogenic differentiation of MSC. This may influence future clinical applications. |
| first_indexed | 2024-12-19T14:15:22Z |
| format | Article |
| id | doaj.art-e0a86fbcf1d2461d96b6f182779f6796 |
| institution | Directory Open Access Journal |
| issn | 2035-8237 2035-8164 |
| language | English |
| last_indexed | 2024-12-19T14:15:22Z |
| publishDate | 2012-12-01 |
| publisher | Open Medical Publishing |
| record_format | Article |
| series | Orthopedic Reviews |
| spelling | doaj.art-e0a86fbcf1d2461d96b6f182779f67962022-12-21T20:18:01ZengOpen Medical PublishingOrthopedic Reviews2035-82372035-81642012-12-0144e36e3610.4081/or.2012.e362365Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cellsFalk Mittag0Eva-Maria Falkenberg1Alexandra Janczyk2Marco Götze3Tino Felka4Wilhelm K. Aicher5Torsten Kluba6Department of Orthopedic Surgery, University Hospital TübingenDepartment of Orthopedic Surgery, University Hospital TübingenDepartment of Orthopedic Surgery, University Hospital TübingenDepartment of Orthopedic Surgery, University Hospital TübingenUniversity Tübingen, Center for Regenerative Biology and Regenerative Medicine TübingenUniversity Tübingen, Center for Regenerative Biology and Regenerative Medicine TübingenDepartment of Orthopedic Surgery, University Hospital TübingenMesenchymal stromal cells (MSC) are differentiation competent cells and may generate, among others, mature osteoblasts or chondrocytes<em> in vitro</em> and <em>in vivo</em>. Laminin-5 and type I collagen are important components of the extracellular matrix. They are involved in a variety of cellular and extracellular activities including cell attachment and osteogenic differentiation of MSC. MSC were isolated and expanded using media conforming good medical practice (GMP)-regulations for medical products. Cells were characterized according to the defined minimal criteria for multipotent MSC. MTT- and BrdU-assays were performed to evaluate protein-dependent (laminin-5, laminin-1, type I collagen) metabolic activity and proliferation of MSC. MSC-attachment assays were performed using protein-coated culture plates. Osteogenic differentiation of MSC was measured by protein-dependant mineralization and expression of osteogenic marker genes (osteopontin, alkaline phophatase, Runx2) after three, seven and 28 days of differentiation. Marker genes were identified using quantitative reverse-transcription polymerase chain reaction. Expansion of MSC in GMP-conforming media yielded vital cells meeting all minimal criteria for MSC. Attachment assay revealed a favorable binding of MSC to laminin-5 and type I collagen at a protein concentration of 1-5 fmol/mL. Compared to plastic, osteogenic differentiation was significantly increased by laminin-5 after 28 days of culture (P<0.04). No significant differences in gene expression patterns were observed. We conclude that laminin-5 and type I collagen promote attachment, but laminin-1 and laminin-5 promote osteogenic differentiation of MSC. This may influence future clinical applications.http://www.pagepress.org/journals/index.php/or/article/view/4467attachment, mesenchymal stromal cells, osteogenic differentiation, type I collagen, laminin-5 |
| spellingShingle | Falk Mittag Eva-Maria Falkenberg Alexandra Janczyk Marco Götze Tino Felka Wilhelm K. Aicher Torsten Kluba Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells Orthopedic Reviews attachment, mesenchymal stromal cells, osteogenic differentiation, type I collagen, laminin-5 |
| title | Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells |
| title_full | Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells |
| title_fullStr | Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells |
| title_full_unstemmed | Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells |
| title_short | Laminin-5 and type I collagen promote adhesion and osteogenic differentiation of animal serum-free expanded human mesenchymal stromal cells |
| title_sort | laminin 5 and type i collagen promote adhesion and osteogenic differentiation of animal serum free expanded human mesenchymal stromal cells |
| topic | attachment, mesenchymal stromal cells, osteogenic differentiation, type I collagen, laminin-5 |
| url | http://www.pagepress.org/journals/index.php/or/article/view/4467 |
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