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Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells

Abubakar Garba UGent, Delphine Acar UGent, Inge Roukaerts, Lowiese Desmarets, Bert Devriendt UGent and Hans Nauwynck UGent (2017) VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY. 191. p.44-50
abstract
Mesenchymal cells are multipotent stromal cells with self-renewal, differentiation and immunomodulatory capabilities. We aimed to develop a co-culture model for differentiating hematopoietic cells on top of immortalized mesenchymal cells for studying interactions between hematopoietic and mesenchymal cells, useful for adequately exploring the therapeutic potential of mesenchymal cells. In this study, we investigated the survival, proliferation and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized porcine bone marrow mesenchymal cells for a period of five weeks. Directly after collection, primary porcine bone marrow mesenchymal cells adhered firmly to the bottom of the culture plates and showed a fibroblast-like appearance, one week after isolation. Upon immortalization, porcine bone marrow mesenchymal cells were continuously proliferating. They were positive for simian virus 40 (SV40) large T antigen and the mesenchymal cell markers CD44 and CD55. Isolated red bone marrow cells were added to these immortalized mesenchymal cells. Five weeks post-seeding, 92 +/- 6% of the red bone marrow hematopoietic cells were still alive and their number increased 3-fold during five weekly subpassages on top of the immortalized mesenchymal cells. The red bone marrow hematopoietic cells were originally small and round; later, the cells increased in size. Some of them became elongated, while others remained round. Tiny dendrites appeared attaching hematopoietic cells to the underlying immortalized mesenchymal cells. Furthermore, weekly differential-quick staining of the cells indicated the presence of monoblasts, monocytes, macrophages and lymphocytes in the co-cultures. At three weeks of co-culture, flow cytometry analysis showed an increased surface expression of CD172a, CD14, CD163, CD169, CD4 and CD8 up to 37 +/- 0.8%, 40 +/- 8%, 41 +/- 4%, 23 +/- 3% and 19 +/- 5% of the hematopoietic cells, respectively. In conclusion, continuous mesenchymal cell cultures were successfully established and characterized and they supported the proliferation of red bone marrow hematopoietic cells, which finally differentiated into monocytic cells and CD4(+) and CD8(+) cells.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Red bone marrow, Hematopoietic cells, Mesenchymal cells, Swine, SHEEP ERYTHROCYTE RECEPTOR, RESPIRATORY SYNDROME VIRUS, STEM-CELLS, PROGENITOR CELLS, STROMAL CELLS, SIALOADHESIN, POPULATIONS, MACROPHAGES, ACTIVATION, EXPRESSION
journal title
VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY
Vet. Immunol. Immunopathol.
volume
191
pages
44 - 50
Web of Science type
Article
Web of Science id
000412254100007
ISSN
0165-2427
DOI
10.1016/j.vetimm.2017.08.002
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
8552143
handle
http://hdl.handle.net/1854/LU-8552143
date created
2018-02-27 12:24:19
date last changed
2018-03-08 15:14:03
@article{8552143,
  abstract     = {Mesenchymal cells are multipotent stromal cells with self-renewal, differentiation and immunomodulatory capabilities. We aimed to develop a co-culture model for differentiating hematopoietic cells on top of immortalized mesenchymal cells for studying interactions between hematopoietic and mesenchymal cells, useful for adequately exploring the therapeutic potential of mesenchymal cells. In this study, we investigated the survival, proliferation and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized porcine bone marrow mesenchymal cells for a period of five weeks. Directly after collection, primary porcine bone marrow mesenchymal cells adhered firmly to the bottom of the culture plates and showed a fibroblast-like appearance, one week after isolation. Upon immortalization, porcine bone marrow mesenchymal cells were continuously proliferating. They were positive for simian virus 40 (SV40) large T antigen and the mesenchymal cell markers CD44 and CD55. Isolated red bone marrow cells were added to these immortalized mesenchymal cells. Five weeks post-seeding, 92 +/- 6\% of the red bone marrow hematopoietic cells were still alive and their number increased 3-fold during five weekly subpassages on top of the immortalized mesenchymal cells. The red bone marrow hematopoietic cells were originally small and round; later, the cells increased in size. Some of them became elongated, while others remained round. Tiny dendrites appeared attaching hematopoietic cells to the underlying immortalized mesenchymal cells. Furthermore, weekly differential-quick staining of the cells indicated the presence of monoblasts, monocytes, macrophages and lymphocytes in the co-cultures. At three weeks of co-culture, flow cytometry analysis showed an increased surface expression of CD172a, CD14, CD163, CD169, CD4 and CD8 up to 37 +/- 0.8\%, 40 +/- 8\%, 41 +/- 4\%, 23 +/- 3\% and 19 +/- 5\% of the hematopoietic cells, respectively. In conclusion, continuous mesenchymal cell cultures were successfully established and characterized and they supported the proliferation of red bone marrow hematopoietic cells, which finally differentiated into monocytic cells and CD4(+) and CD8(+) cells.},
  author       = {Garba, Abubakar and Acar, Delphine and Roukaerts, Inge and Desmarets, Lowiese and Devriendt, Bert and Nauwynck, Hans},
  issn         = {0165-2427},
  journal      = {VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY},
  keyword      = {Red bone marrow,Hematopoietic cells,Mesenchymal cells,Swine,SHEEP ERYTHROCYTE RECEPTOR,RESPIRATORY SYNDROME VIRUS,STEM-CELLS,PROGENITOR CELLS,STROMAL CELLS,SIALOADHESIN,POPULATIONS,MACROPHAGES,ACTIVATION,EXPRESSION},
  language     = {eng},
  pages        = {44--50},
  title        = {Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells},
  url          = {http://dx.doi.org/10.1016/j.vetimm.2017.08.002},
  volume       = {191},
  year         = {2017},
}

Chicago
Garba, Abubakar, Delphine Acar, Inge Roukaerts, Lowiese Desmarets, Bert Devriendt, and Hans Nauwynck. 2017. “Long-term Culture and Differentiation of Porcine Red Bone Marrow Hematopoietic Cells Co-cultured with Immortalized Mesenchymal Cells.” Veterinary Immunology and Immunopathology 191: 44–50.
APA
Garba, A., Acar, D., Roukaerts, I., Desmarets, L., Devriendt, B., & Nauwynck, H. (2017). Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY, 191, 44–50.
Vancouver
1.
Garba A, Acar D, Roukaerts I, Desmarets L, Devriendt B, Nauwynck H. Long-term culture and differentiation of porcine red bone marrow hematopoietic cells co-cultured with immortalized mesenchymal cells. VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY. 2017;191:44–50.
MLA
Garba, Abubakar, Delphine Acar, Inge Roukaerts, et al. “Long-term Culture and Differentiation of Porcine Red Bone Marrow Hematopoietic Cells Co-cultured with Immortalized Mesenchymal Cells.” VETERINARY IMMUNOLOGY AND IMMUNOPATHOLOGY 191 (2017): 44–50. Print.