Research Article Attachment, Growth, and Detachment of Human Mesenchymal Stem Cells in a Chemically Defined Medium Denise Salzig, 1 Jasmin Leber, 1 Katharina Merkewitz, 1 Michaela C. Lange, 1 Natascha Köster, 1 and Peter Czermak 1,2,3,4 1 Institute of Bioprocess Engineering and Pharmaceutical Technology, University of Applied Sciences Mittelhessen, 35390 Giessen, Germany 2 Faculty of Biology and Chemistry, Justus Liebig University, 35390 Giessen, Germany 3 Project Group Bioresources, Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), 35394 Giessen, Germany 4 Department of Chemical Engineering, Kansas State University, Manhattan, KS 66506, USA Correspondence should be addressed to Denise Salzig; denise.salzig@lse.thm.de Received 8 October 2015; Revised 18 January 2016; Accepted 26 January 2016 Academic Editor: Tao-Sheng Li Copyright © 2016 Denise Salzig et al. Tis is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Te manufacture of human mesenchymal stem cells (hMSCs) for clinical applications requires an appropriate growth surface and an optimized, preferably chemically defned 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 fbers. Cell spreading was promoted by coating the growth surface with collagen type IV and fbronectin. Te 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 efect. All three cell types retained their trilineage diferentiation capability in CDM and were detached by several enzymes (but not collagenase in the case of hMSC-TERT cells). Te medium and coating did not afect detachment efciency but infuenced cell survival afer detachment. CDM combined with cell-specifc surface coatings and/or FGF-2 supplements is therefore as efective as serum-containing medium for the manufacture of diferent hMSC types. 1. Introduction Human mesenchymal stem/stromal cells (hMSCs) are ofen used for cell therapy because they ofer many advantageous characteristics [1]. Before therapeutic use, hMSCs must be expanded to produce the number of cells needed per patient and per dose (at least 1-2 × 10 6 hMSCs per kg) [2]. Te growth of hMSCs is anchorage-dependent, and the interactions among the growth surface, cells, and surrounding medium are therefore important for the manufacture of suitable numbers of healthy cells. Cell adhesion is necessary for hMSC expansion and is driven by both nonspecifc and specifc interactions. Rounded cells in suspension initially attach to the surface due to complementary electrostatic/ionic forces and the growth surface then interacts with cell surface integrins, the principal receptors mediating cell-matrix adhesion [3]. Integrin activation results in the formation of heterodimers, which initiate signaling cascades that activate downstream genes and ultimately regulate cell morphology and behavior. Te cell fattens and spreads due to the activation of protein kinase C (PKC) and the subsequent accumulation of focal adhesion kinase (FAK) and actin flaments at the leading edges of the cells. Te completion of cell spreading and strong adhesion to the surface, which is required for proliferation, is characterized by the inactivation of PKC and the cross- linking of actin to defned intracellular stress fbers along with FAK located at the focal adhesion sites. Te actin forms a stable cytoskeleton, which maintains the cell in its adherent spread state [4, 5]. Hindawi Publishing Corporation Stem Cells International Volume 2016, Article ID 5246584, 10 pages http://dx.doi.org/10.1155/2016/5246584