Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium

The manufacture of human mesenchymal stem cells (hMSCs) for clinical applications requires an appropriate growth surface and an optimized, preferably chemically defined medium (CDM) for expansion. We investigated a new protein/peptide-free CDM that supports the adhesion, growth, and detachment of an...

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Main Authors: Denise Salzig, Jasmin Leber, Katharina Merkewitz, Michaela C. Lange, Natascha Köster, Peter Czermak
Format: Article
Language:English
Published: Wiley 2016-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2016/5246584
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author Denise Salzig
Jasmin Leber
Katharina Merkewitz
Michaela C. Lange
Natascha Köster
Peter Czermak
author_facet Denise Salzig
Jasmin Leber
Katharina Merkewitz
Michaela C. Lange
Natascha Köster
Peter Czermak
author_sort Denise Salzig
collection DOAJ
description The manufacture of human mesenchymal stem cells (hMSCs) for clinical applications requires an appropriate growth surface and an optimized, preferably chemically defined medium (CDM) for expansion. We investigated a new protein/peptide-free CDM that supports the adhesion, growth, and detachment of an immortalized hMSC line (hMSC-TERT) as well as primary cells derived from bone marrow (bm-hMSCs) and adipose tissue (ad-hMSCs). We observed the rapid attachment and spreading of hMSC-TERT cells and ad-hMSCs in CDM concomitant with the expression of integrin and actin fibers. Cell spreading was promoted by coating the growth surface with collagen type IV and fibronectin. The growth of hMSC-TERT cells was similar in CDM and serum-containing medium whereas the lag phase of bm-hMSCs was prolonged in CDM. FGF-2 or surface coating with collagen type IV promoted the growth of bm-hMSCs, but laminin had no effect. All three cell types retained their trilineage differentiation capability in CDM and were detached by several enzymes (but not collagenase in the case of hMSC-TERT cells). The medium and coating did not affect detachment efficiency but influenced cell survival after detachment. CDM combined with cell-specific surface coatings and/or FGF-2 supplements is therefore as effective as serum-containing medium for the manufacture of different hMSC types.
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spelling doaj-art-e3be4c84ae384874aa0f8bf8c6b42de12025-08-20T03:26:31ZengWileyStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/52465845246584Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined MediumDenise Salzig0Jasmin Leber1Katharina Merkewitz2Michaela C. Lange3Natascha Köster4Peter Czermak5Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, GermanyInstitute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, GermanyThe manufacture of human mesenchymal stem cells (hMSCs) for clinical applications requires an appropriate growth surface and an optimized, preferably chemically defined medium (CDM) for expansion. We investigated a new protein/peptide-free CDM that supports the adhesion, growth, and detachment of an immortalized hMSC line (hMSC-TERT) as well as primary cells derived from bone marrow (bm-hMSCs) and adipose tissue (ad-hMSCs). We observed the rapid attachment and spreading of hMSC-TERT cells and ad-hMSCs in CDM concomitant with the expression of integrin and actin fibers. Cell spreading was promoted by coating the growth surface with collagen type IV and fibronectin. The growth of hMSC-TERT cells was similar in CDM and serum-containing medium whereas the lag phase of bm-hMSCs was prolonged in CDM. FGF-2 or surface coating with collagen type IV promoted the growth of bm-hMSCs, but laminin had no effect. All three cell types retained their trilineage differentiation capability in CDM and were detached by several enzymes (but not collagenase in the case of hMSC-TERT cells). The medium and coating did not affect detachment efficiency but influenced cell survival after detachment. CDM combined with cell-specific surface coatings and/or FGF-2 supplements is therefore as effective as serum-containing medium for the manufacture of different hMSC types.http://dx.doi.org/10.1155/2016/5246584
spellingShingle Denise Salzig
Jasmin Leber
Katharina Merkewitz
Michaela C. Lange
Natascha Köster
Peter Czermak
Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium
Stem Cells International
title Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium
title_full Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium
title_fullStr Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium
title_full_unstemmed Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium
title_short Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium
title_sort attachment growth and detachment of human mesenchymal stem cells in a chemically defined medium
url http://dx.doi.org/10.1155/2016/5246584
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