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Optimization of the isolation, culture, and characterization of equine umbilical cord blood mesenchymal stromal cells

(2011) TISSUE ENGINEERING PART C-METHODS. 17(11). p.1061-1070
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Abstract
Mesenchymal stromal cells (MSC) represent a promising population for supporting new clinical concepts in cellular therapy. A wide diversity of isolation procedures for MSC from umbilical cord blood (UCB) has been described for humans. In contrast, a few data are available in horses. In the current study, a sedimentation method using hydroxyethyl starch and a method based on the lysis of red blood cells using ammonium chloride (NH(4)Cl) were compared with two density gradient separation methods (Ficoll-Paque and Percoll). Adherent cell colonies could be established using all four isolation methods. The mononuclear cell recovery after Percoll separation, however, resulted in significantly more putative MSC colonies; and, therefore, this isolation method was used for all further experiments. Culture conditions such as cell density and medium or serum coating of the wells did not significantly affect putative MSC recovery. Isolated MSC using Percoll were subsequently differentiated toward the osteogenic, chondrogenic, and adipogenic lineage. In addition, MSC were phenotyped by multicolor flow cytometry based on their expression of different cell protein markers. Cultured MSC were CD29, CD44, and CD90-positive and CD79 alpha, Macrophage/Monocyte and MHC II-negative. In conclusion, this study reports optimized protocols to isolate, culture, and characterize solid equine MSC from UCB.
Keywords
BONE-MARROW, STEM-CELLS, DENSITY-SEPARATION, PROGENITOR CELLS, UNRELATED TRANSPLANTATION, IN-VITRO, CRYOPRESERVATION, DIFFERENTIATION, COLLECTION, BANKING

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Chicago
De Schauwer, Catharina, Evelyne Meyer, Pieter Cornillie, Sarne De Vliegher, Gerlinde Van de Walle, Maarten Hoogewijs, Heidi Declercq, et al. 2011. “Optimization of the Isolation, Culture, and Characterization of Equine Umbilical Cord Blood Mesenchymal Stromal Cells.” Tissue Engineering Part C-methods 17 (11): 1061–1070.
APA
De Schauwer, C., Meyer, E., Cornillie, P., De Vliegher, S., Van de Walle, G., Hoogewijs, M., Declercq, H., et al. (2011). Optimization of the isolation, culture, and characterization of equine umbilical cord blood mesenchymal stromal cells. TISSUE ENGINEERING PART C-METHODS, 17(11), 1061–1070.
Vancouver
1.
De Schauwer C, Meyer E, Cornillie P, De Vliegher S, Van de Walle G, Hoogewijs M, et al. Optimization of the isolation, culture, and characterization of equine umbilical cord blood mesenchymal stromal cells. TISSUE ENGINEERING PART C-METHODS. 2011;17(11):1061–70.
MLA
De Schauwer, Catharina, Evelyne Meyer, Pieter Cornillie, et al. “Optimization of the Isolation, Culture, and Characterization of Equine Umbilical Cord Blood Mesenchymal Stromal Cells.” TISSUE ENGINEERING PART C-METHODS 17.11 (2011): 1061–1070. Print.
@article{1954856,
  abstract     = {Mesenchymal stromal cells (MSC) represent a promising population for supporting new clinical concepts in cellular therapy. A wide diversity of isolation procedures for MSC from umbilical cord blood (UCB) has been described for humans. In contrast, a few data are available in horses. In the current study, a sedimentation method using hydroxyethyl starch and a method based on the lysis of red blood cells using ammonium chloride (NH(4)Cl) were compared with two density gradient separation methods (Ficoll-Paque and Percoll). Adherent cell colonies could be established using all four isolation methods. The mononuclear cell recovery after Percoll separation, however, resulted in significantly more putative MSC colonies; and, therefore, this isolation method was used for all further experiments. Culture conditions such as cell density and medium or serum coating of the wells did not significantly affect putative MSC recovery. Isolated MSC using Percoll were subsequently differentiated toward the osteogenic, chondrogenic, and adipogenic lineage. In addition, MSC were phenotyped by multicolor flow cytometry based on their expression of different cell protein markers. Cultured MSC were CD29, CD44, and CD90-positive and CD79 alpha, Macrophage/Monocyte and MHC II-negative. In conclusion, this study reports optimized protocols to isolate, culture, and characterize solid equine MSC from UCB.},
  author       = {De Schauwer, Catharina and Meyer, Evelyne and Cornillie, Pieter and De Vliegher, Sarne and Van de Walle, Gerlinde and Hoogewijs, Maarten and Declercq, Heidi and Govaere, Jan and Demeyere, Kristel and Cornelissen, Maria and Van Soom, Ann},
  issn         = {1937-3384},
  journal      = {TISSUE ENGINEERING PART C-METHODS},
  keyword      = {BONE-MARROW,STEM-CELLS,DENSITY-SEPARATION,PROGENITOR CELLS,UNRELATED TRANSPLANTATION,IN-VITRO,CRYOPRESERVATION,DIFFERENTIATION,COLLECTION,BANKING},
  language     = {eng},
  number       = {11},
  pages        = {1061--1070},
  title        = {Optimization of the isolation, culture, and characterization of equine umbilical cord blood mesenchymal stromal cells},
  url          = {http://dx.doi.org/10.1089/ten.tec.2011.0052},
  volume       = {17},
  year         = {2011},
}

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