title: Human AB serum and thrombin-activated platelet-rich plasma are suitable alternatives to fetal calf serum for the expansion of mesenchymal stem cells from adipose tissue.
abstract: MSCs are currently in focus regarding their clinical potential in cell therapy and tissue engineering. However, most isolation and expansion protocols for clinical-scale production of MSCs use fetal calf serum (FCS) as a supplement, which poses a potential risk for infections as well as immunological reactions. To find a suitable FCS substitute, we investigated the effects of pooled human AB serum (AB-HS) and thrombin-activated platelet-rich plasma (tPRP) on adipose tissue MSCs (AT-MSCs) with FCS as the standard control medium. AT-MSCs of 10 donors were cultured under three different conditions: (a) 10% FCS, (b) 10% AB-HS, and (c) 10% tPRP. Colony-forming units, cumulative population doubling rates, and differentiation capacity toward the adipogenic and osteogenic lineages were assessed, along with immunophenotype. We demonstrated that AB-HS and tPRP provide a significantly higher proliferative effect on AT-MSCs than does FCS. In the first six passages, AB-HS and tPRP MSCs exhibited a fold expansion of 66.6 +/- 15.7 and 68.1 +/- 6.7, respectively, compared with 24.4 +/- 0.7 for FCS. Differentiation capacity was preserved throughout long-term culture. Immunophenotype was characteristic for MSCs and comparable for all culture conditions with the exception of a distinct CD45-/CD14-positive side population for AB-HS and tPRP that tended to diminish with prolonged culture. We showed that pooled human AB serum and thrombin-activated platelet-rich plasma are alternatives to FCS for AT-MSCs. These human sources are better characterized regarding potential infectious threats, while providing a higher proliferation rate and retaining differentiation capacity and mesenchymal stem cell marker expression throughout long-term culture. Disclosure of potential conflicts of interest is found at the end of this article.
authors.affiliation: Institute of Transfusion Medicine and Immunology, German Red Cross Blood Service of Baden-Württemberg Hessen, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany.
authors.forename: Asli
authors.initials: A
authors.lastname: Kocaoemer
authors.forename: Susanne
authors.initials: S
authors.lastname: Kern
authors.forename: Harald
authors.initials: H
authors.lastname: Klüter
authors.forename: Karen
authors.initials: K
authors.lastname: Bieback
mesh_terms: D000017:ABO Blood-Group System / Q000097:blood*; D000273:Adipose Tissue / Q000166:cytology* / Q000187:drug effects; D000328:Adult; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects; D048708:Cell Growth Processes / Q000187:drug effects; D002469:Cell Separation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D016207:Cytokines / Q000378:metabolism; D005260:Female; D006801:Humans; D017493:Leukocyte Common Antigens / Q000378:metabolism; D018950:Lipopolysaccharide Receptors / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D008875:Middle Aged; D015539:Platelet Activation / Q000187:drug effects*; D053657:Platelet-Rich Plasma / Q000378:metabolism*; D044967:Serum / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology*
chemical_list: D000017:ABO Blood-Group System; D015415:Biomarkers; D016207:Cytokines; D018950:Lipopolysaccharide Receptors; D017493:Leukocyte Common Antigens; D013917:Thrombin