Impact of individual platelet lysates on isolation and growth of human mesenchymal stromal cells.

BACKGROUND AIMS Culture medium for mesenchymal stromal cells (MSC) is frequently supplemented with fetal calf serum (FCS). FCS can induce xenogeneic immune reactions, transmit bovine pathogens and has a high lot-to-lot variability that hampers reproducibility of results. Several studies have demonstrated that pooled human platelet lysate (HPL) provides an attractive alternative for FCS. However, little is known about the variation between different platelet lysates. METHODS We compared activities of individual HPL on initial fibroblastoid colony-forming units (CFU-F), proliferation, in vitro differentiation and long-term culture. These data were correlated with chemokine profiles of HPL. RESULTS Isolation of MSC with either HPL or FCS resulted in similar CFU-F frequency, colony morphology, immunophenotype and adipogenic differentiation potential. Osteogenic differentiation was even more pronounced in HPL than FCS. There were significant differences in MSC proliferation with different HPL, but it was always higher in comparison with FCS. Cell growth correlated with the concentration of platelet-derived growth factor (PDGF) and there was a moderate association with platelet counts. All HPL facilitated expansion for more than 20 population doublings. CONCLUSIONS Taken together, reliable long-term expansion was possible with all HPL, although there was some variation in platelet lysates of individual units. Therefore the use of donor recipient-matched or autologous HPL is feasible for therapeutic MSC products.

[1]  Karl Kashofer,et al.  Human platelet lysate can replace fetal bovine serum for clinical‐scale expansion of functional mesenchymal stromal cells , 2007, Transfusion.

[2]  F. Marini,et al.  Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement. , 2005, Cytotherapy.

[3]  A. Ho,et al.  Mesenchymal Stem Cell Preparations—Comparing Apples and Oranges , 2007, Stem Cell Reviews.

[4]  Y. Verma,et al.  Mesenchymal stem cell-based therapy: a new paradigm in regenerative medicine , 2009, Journal of cellular and molecular medicine.

[5]  A. Kiani,et al.  Treatment of refractory acute GVHD with third-party MSC expanded in platelet lysate-containing medium , 2009, Bone Marrow Transplantation.

[6]  C Roskelley,et al.  A biomarker that identifies senescent human cells in culture and in aging skin in vivo. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[7]  H. Zhang,et al.  Mesenchymal stem cells and cardiac repair , 2008, Journal of cellular and molecular medicine.

[8]  C. Verfaillie,et al.  Erratum:Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood. 2001; 98:2615-2625 , 2009 .

[9]  T. Mosmann Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[10]  V. Beneš,et al.  Replicative Senescence of Mesenchymal Stem Cells: A Continuous and Organized Process , 2008, PloS one.

[11]  E. Andreu,et al.  Comparison of ex vivo expansion culture conditions of mesenchymal stem cells for human cell therapy , 2009, Transfusion.

[12]  Peter Harrison,et al.  Platelet a-granules , 1993 .

[13]  A. Ho,et al.  REVIEW Heterogeneity of mesenchymal stromal cell preparations , 2008 .

[14]  M. Owen Marrow stromal stem cells , 1988, Journal of Cell Science.

[15]  G. Cagney,et al.  Characterization of the proteins released from activated platelets leads to localization of novel platelet proteins in human atherosclerotic lesions. , 2004, Blood.

[16]  A. Cometa,et al.  Generation of mesenchymal stromal cells in the presence of platelet lysate: a phenotypic and functional comparison of umbilical cord blood- and bone marrow-derived progenitors , 2009, Haematologica.

[17]  B. Korn,et al.  DNA methylation pattern changes upon long-term culture and aging of human mesenchymal stromal cells , 2010, Aging cell.

[18]  D. Prockop,et al.  Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. , 2006, Cytotherapy.

[19]  W. Wagner,et al.  Standardized isolation of human mesenchymal stromal cells with red blood cell lysis. , 2011, Methods in molecular biology.

[20]  P. Robey,et al.  EFFECT OF SERUM ON HUMAN BONE MARROW STROMAL CELLS: EX VIVO EXPANSION AND IN VIVO BONE FORMATION , 2000, Transplantation.

[21]  C. Choong,et al.  PDGF, TGF-beta, and FGF signaling is important for differentiation and growth of mesenchymal stem cells (MSCs): transcriptional profiling can identify markers and signaling pathways important in differentiation of MSCs into adipogenic, chondrogenic, and osteogenic lineages. , 2008, Blood.

[22]  J. Krieger,et al.  TRANSPLANTATION AND CELLULAR ENGINEERING: Adipose tissue mesenchymal stem cell expansion in animal serum‐free medium supplemented with autologous human platelet lysate , 2009, Transfusion.

[23]  A. Zander,et al.  Autologous serum for isolation and expansion of human mesenchymal stem cells for clinical use. , 2004, Experimental hematology.

[24]  X. Holy,et al.  Platelet lysates promote mesenchymal stem cell expansion: A safety substitute for animal serum in cell‐based therapy applications , 2005, Journal of cellular physiology.

[25]  A. Ho,et al.  The heterogeneity of human mesenchymal stem cell preparations--evidence from simultaneous analysis of proteomes and transcriptomes. , 2006, Experimental hematology.

[26]  Alan K. Smith,et al.  Alternatives to animal sera for human bone marrow cell expansion: human serum and serum-free media. , 1998, Journal of hematotherapy.

[27]  Wolfgang Wagner,et al.  Replicative senescence-associated gene expression changes in mesenchymal stromal cells are similar under different culture conditions , 2010, Haematologica.

[28]  D. G. Halme,et al.  FDA regulation of stem-cell-based therapies. , 2006, The New England journal of medicine.

[29]  H. Klüter,et al.  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 , 2007, Stem cells.

[30]  J. Laine,et al.  N‐Glycolylneuraminic Acid Xenoantigen Contamination of Human Embryonic and Mesenchymal Stem Cells Is Substantially Reversible , 2007, Stem cells.

[31]  Paul J. Harrison,et al.  Platelet alpha-granules. , 1993, Blood reviews.

[32]  D. Strunk,et al.  Human Alternatives to Fetal Bovine Serum for the Expansion of Mesenchymal Stromal Cells from Bone Marrow , 2009, Stem cells.

[33]  A. Ho,et al.  Isolation of human mesenchymal stromal cells is more efficient by red blood cell lysis. , 2008, Cytotherapy.

[34]  A. Ho,et al.  Heterogeneity of mesenchymal stromal cell preparations. , 2008, Cytotherapy.

[35]  R. G. Allen,et al.  Relationship between donor age and the replicative lifespan of human cells in culture: a reevaluation. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[36]  J. Blanco,et al.  Optimization of mesenchymal stem cell expansion procedures by cell separation and culture conditions modification. , 2008, Experimental hematology.

[37]  M. Pittenger,et al.  Multilineage potential of adult human mesenchymal stem cells. , 1999, Science.

[38]  William Stafford Noble,et al.  Analysis of strain and regional variation in gene expression in mouse brain , 2001, Genome Biology.

[39]  M. Introna,et al.  Human platelet lysate allows expansion and clinical grade production of mesenchymal stromal cells from small samples of bone marrow aspirates or marrow filter washouts , 2007, Bone Marrow Transplantation.

[40]  O. Ringdén,et al.  No alloantibodies against mesenchymal stromal cells, but presence of anti-fetal calf serum antibodies, after transplantation in allogeneic hematopoietic stem cell recipients , 2007, Haematologica.

[41]  R. Handgretinger,et al.  Animal serum-free culture conditions for isolation and expansion of multipotent mesenchymal stromal cells from human BM. , 2006, Cytotherapy.

[42]  C. Verfaillie,et al.  Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. , 2001, Blood.

[43]  R Cancedda,et al.  Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. , 2000, Journal of cell science.

[44]  M. Yamaguchi,et al.  Bone marrow stromal cells prepared using AB serum and bFGF for hematopoietic stem cells expansion , 2002, Transfusion.

[45]  M. Krampera,et al.  Mesenchymal stem cells for clinical application , 2010, Vox sanguinis.

[46]  A. Ho,et al.  Molecular and Secretory Profiles of Human Mesenchymal Stromal Cells and Their Abilities to Maintain Primitive Hematopoietic Progenitors , 2007, Stem cells.

[47]  D. Prockop,et al.  The Wnt Signaling Inhibitor Dickkopf-1 Is Required for Reentry into the Cell Cycle of Human Adult Stem Cells from Bone Marrow* , 2003, Journal of Biological Chemistry.

[48]  W. Ansorge,et al.  Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. , 2005, Experimental hematology.

[49]  Philippe Bierling,et al.  Osteoblastic differentiation of human mesenchymal stem cells with platelet lysate. , 2010, Biomaterials.

[50]  H. Klüter,et al.  Critical Parameters for the Isolation of Mesenchymal Stem Cells from Umbilical Cord Blood , 2004, Stem cells.

[51]  A. Reinisch,et al.  Rapid large-scale expansion of functional mesenchymal stem cells from unmanipulated bone marrow without animal serum. , 2008, Tissue engineering. Part C, Methods.