Homing Genes Expression in Fucosyltransferase VI-Treated Umbilical Cord Blood CD133+ Cells which Expanded on Protein-Coated Nanoscaffolds
Umbilical cord blood (UCB)-derived hematopoietic stem cells (HSCs) are considered because of their self-renewing, differentiating, proliferating, and readily available properties. Moreover, HSCsʼ homing to the hematopoietic microenvironment is an important step in their transplantation process. But low content of progenitor cells in one unit of UCB and defect in the bone marrow (BM) homing limit their applications. Hence, we decided to correct this deficiency with ex vivo incubation of CD133+ cells using fucosyltransferase VI and GDP-fucose. Then C-X-C chemokines receptor-4 (CXCR4), very late activation antigen-4 (VLA4), very late activation antigen-5 (VLA5), lymphocyte function-associated antigen-1 (LFA-1), and E-cadherin (E-cad) genes expressions were investigated with the goal of homing evaluation. The purity of MACS isolated CD133+ cells and confirmation of fucosylation were done by flow cytometry, and the viability of cells seeded on protein-coated poly l-lactic acid (PLLA) scaffold was proven via MTT assay. Scanning electron microscopy (SEM), CFU assays, and expression assays of CXCR4, VLA4, VLA5, LFA-1 and E-cad by real-time PCR were performed, too. Flow cytometry data showed that isolated cells were suitable for fucosyltransferase VI (FT-VI) incubation and expansion on nanoscaffolds. MTT, CFU assays, and SEM micrographs demonstrated fibronectin (FN)–collagen–selectin (FCS)-coated scaffold serve as best environment for viability, clonogenicity, and cell attachment. High levels of homing genes expression were also observed in cells seeded on FCS-coated scaffolds. Also, CXCR4 flow cytometry analysis confirmed real-time data. FCS-PLLA scaffolds provided optimal conditions for viability of FT-VI-treated CD133+ cells, and clonogenicity with the goal of improving homing following UCB-HSCs transplantation.
KeywordsCord blood stem cells Fucosyltransferase CXCR4 VLA4 VLA5
The authors would like to thank Bonyakhteh Research Center for providing laboratory facilities. The authors declare that there is no conflict of interest regarding the publication of this article.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 6.Connor, N. S., Aubin, J. E., & Sodek, J. (1983). Independent expression of type I collagen and fibronectin by normal fibroblast-like cells. Journal of Cell Science, 63, 233–244.Google Scholar
- 10.Eskandari, F., Allahverdi, A., Nasiri, H., Azad, M., Kalantari, N., Soleimani, M., & Zare-Zardini, H. (2015). Nanofiber expansion of umbilical cord blood hematopoietic stem cells. Iranian Journal of Pediatric Hematology and Oncology, 5(4), 170–178.Google Scholar
- 14.Greenberg, A. W., Kerr, W. G., & Hammer, D. A. (2000). Relationship between selectin-mediated rolling of hematopoietic stem and progenitor cells and progression in hematopoietic development. Blood, 95(2), 478–486.Google Scholar
- 16.Islami M, Mortazavi Y, Nadri S, Soleimani M (2017) In vitro expansion of CD 133+ cells derived from umbilical cord blood in poly-L-lactic acid (PLLA) scaffold coated with fibronectin and collagen. Artificial Cells, Nanomedicine, and Biotechnology. https://doi.org/10.1080/21691401.2017.1358733 Google Scholar
- 20.Kollet, O., Spiegel, A., Peled, A., Petit, I., Byk, T., Hershkoviz, R., … Lapidot, T. (2001). Rapid and efficient homing of human CD34(+)CD38(-/low)CXCR4(+) stem and progenitor cells to the bone marrow and spleen of NOD/SCID and NOD/SCID/B2m(null) mice. Blood, 97(10), 3283–3291.CrossRefGoogle Scholar
- 27.Papayannopoulou, T., Craddock, C., Nakamoto, B., Priestley, G. V., & Wolf, N. S. (1995). The VLA4/VCAM-1 adhesion pathway defines contrasting mechanisms of lodgement of transplanted murine hemopoietic progenitors between bone marrow and spleen. Proceedings of the National Academy of Sciences, 92(21), 9647–9651.CrossRefGoogle Scholar
- 28.Peled, A., Kollet, O., Ponomaryov, T., Petit, I., Franitza, S., Grabovsky, V., … Lapidot, T. (2000). The chemokine SDF-1 activates the integrins LFA-1, VLA-4, and VLA-5 on immature human CD34(+) cells: Role in transendothelial/stromal migration and engraftment of NOD/SCID mice. Blood, 95(11), 3289–3296.Google Scholar
- 31.Riedl, E., Stockl, J., Majdic, O., Scheinecker, C., Knapp, W., & Strobl, H. (2000). Ligation of E-cadherin on in vitro-generated immature Langerhans-type dendritic cells inhibits their maturation. Blood, 96(13), 4276–4284.Google Scholar
- 32.Sabaghi, F., Shamsasenjan, K., Movasaghpour, A. A., Amirizadeh, N., Nikougoftar, M., & Bagheri, N. (2016). Evaluation of human cord blood CD34+ hematopoietic stem cell differentiation to megakaryocyte on aminated PES nanofiber scaffold compare to 2-D culture system. Artificial Cells, Nanomedicine, and Biotechnology, 44(4), 1062–1068.Google Scholar