SSEA-4 Antigen Is Expressed on Rabbit Lymphocyte Subsets
Abstract
:1. Introduction
2. Results
2.1. Flow-Cytometric Phenotyping of SSEA-4+ Cells
2.2. Magnetic-Activated Cell Sorting (MACS) of SSEA-4+ Cells
2.3. Real-Time (qPCR) Analysis of SSEA-4 Sorted Cells
3. Discussion
4. Materials and Methods
4.1. Animals
4.2. Experimental Design
4.3. Flow Cytometry
4.4. Magnetic-Activated Cell Sorting of SSEA-4+ Cells
4.5. Real-Time PCR (qPCR)
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
Confocal Microscopy
Appendix B
References
- Kannagi, R.; Cochran, N.A.; Ishigami, F.; Hakomori, S.; Andrews, P.W.; Knowles, B.B.; Solter, D. Stage-specific embryonic antigens (SSEA-3 and -4) are epitopes of a unique globo—Series ganglioside isolated from human teratocarcinoma cells. EMBO J. 1983, 2, 2355–2361. [Google Scholar] [CrossRef] [PubMed]
- Muramatsu, T.; Muramatsu, H. Carbohydrate antigens expressed on stem cells and early embryonic cells. Glycoconj. J. 2004, 21, 41–45. [Google Scholar] [CrossRef] [PubMed]
- Wright, A.J.; Andrews, P.W. Surface marker antigens in the characterization of human embryonic stem cells. Stem Cell Res. 2009, 3, 3–11. [Google Scholar] [CrossRef] [Green Version]
- Suila, H.; Pitkanen, V.; Hirvonen, T.; Heiskanen, A.; Anderson, H.; Laitinen, A.; Natunen, S.; Miller-Podraza, H.; Satomaa, T.; Natunen, J.; et al. Are globoseries glycosphingolipids SSEA-3 and-4 markers for stem cells derived from human umbilical cord blood? J. Mol. Cell Biol. 2011, 3, 99–107. [Google Scholar] [CrossRef]
- Bhartiya, D.; Shaikh, A.; Nagvenkar, P.; Kasiviswanathan, S.; Pethe, P.; Pawani, H.; Mohanty, S.; Rao, S.G.A.; Zaveri, K.; Hinduja, I. Very Small Embryonic-Like Stem Cells with Maximum Regenerative Potential Get Discarded During Cord Blood Banking and Bone Marrow Processing for Autologous Stem Cell Therapy. Stem Cells Dev. 2012, 21, 1–6. [Google Scholar] [CrossRef]
- Gang, E.J.; Bosnakovski, D.; Figueiredo, C.A.; Visser, J.W.; Perlingeiro, R.C.R. SSEA-4 identifies mesenchymal stem cells from bone marrow. Blood 2007, 109, 1743–1751. [Google Scholar] [CrossRef] [PubMed]
- Sandstedt, J.; Jonsson, M.; Vukusic, K.; Dellgren, G.; Lindahl, A.; Jeppsson, A.; Asp, J. SSEA-4+CD34-Cells in the Adult Human Heart Show the Molecular Characteristics of a Novel Cardiomyocyte Progenitor Population. Cells Tissues Organs 2014, 199, 103–116. [Google Scholar] [CrossRef]
- Afrikanova, I.; Kayali, A.; Lopez, A.; Hayek, A. Is Stage-Specific Embryonic Antigen 4 a Marker for Human Ductal Stem/Progenitor Cells? Biores. Open Access 2012, 1, 184–191. [Google Scholar] [CrossRef]
- He, H.P.; Nagamura-Inoue, T.; Tsunoda, H.; Yuzawa, M.; Yamamoto, Y.; Yorozu, P.; Agata, H.; Tojo, A. Stage-Specific Embryonic Antigen 4 in Wharton’s Jelly-Derived Mesenchymal Stem Cells Is Not a Marker for Proliferation and Multipotency. Tissue Eng. Part A 2014, 20, 1314–1324. [Google Scholar] [CrossRef]
- Kuroda, Y.; Kitada, M.; Wakao, S.; Nishikawa, K.; Tanimura, Y.; Makinoshima, H.; Goda, M.; Akashi, H.; Inutsuka, A.; Niwa, A.; et al. Unique multipotent cells in adult human mesenchymal cell populations. Proc. Natl. Acad. Sci. USA 2010, 107, 8639–8643. [Google Scholar] [CrossRef] [Green Version]
- Wakao, S.; Kitada, M.; Kuroda, Y.; Dezawa, M. Isolation of Adult Human Pluripotent Stem Cells from Mesenchymal Cell Populations and Their Application to Liver Damages. In Liver Stem Cells: Methods and Protocols; Methods in Molecular Biology; Ochiya, T., Ed.; Humana Press Inc.: Totowa, NJ, USA, 2012; Volume 826, pp. 89–102. [Google Scholar]
- Brimble, S.N.; Sherrer, E.S.; Uhl, E.W.; Wang, E.; Kelly, S.; Merrill, A.H.; Robins, A.J.; Schulz, T.C. The cell surface glycosphingolipids SSEA-3 and SSEA-4 are not essential for human ESC pluripotency. Stem Cells 2007, 25, 54–62. [Google Scholar] [CrossRef]
- Yamashita, T.; Wada, R.; Sasaki, T.; Deng, C.X.; Bierfreund, U.; Sandhoff, K.; Proia, R.L. A vital role for glycosphingolipid synthesis during development and differentiation. Proc. Natl. Acad. Sci. USA 1999, 96, 9142–9147. [Google Scholar] [CrossRef] [Green Version]
- Dodd, J.; Solter, D.; Jessell, T.M. Monoclonal antibodies against carbohydrate differentiation antigens identify subsets of primary sensory neurones. Nature 1984, 311, 469–472. [Google Scholar] [CrossRef]
- Truong, T.T.; Huynh, K.; Nakatsu, M.N.; Deng, S.X. SSEA4 Is a Potential Negative Marker for the Enrichment of Human Corneal Epithelial Stem/Progenitor Cells. Investig. Ophthalmol. Vis. Sci. 2011, 52, 6315–6320. [Google Scholar] [CrossRef]
- Mueller, T.; Eildermann, K.; Dhir, R.; Schlatt, S.; Behr, R. Glycan stem-cell markers are specifically expressed by spermatogonia in the adult non-human primate testis. Hum. Reprod. 2008, 23, 2292–2298. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Rozemuller, H.; Prins, H.J.; Naaijkens, B.; Staal, J.; Buhring, H.J.; Martens, A.C. Prospective Isolation of Mesenchymal Stem Cells from Multiple Mammalian Species Using Cross-Reacting Anti-Human Monoclonal Antibodies. Stem Cells Dev. 2010, 19, 1911–1921. [Google Scholar] [CrossRef] [PubMed]
- Ding, Z.; Huang, H. Mesenchymal stem cells in rabbit meniscus and bone marrow exhibit a similar feature but a heterogeneous multi-differentiation potential: Superiority of meniscus as a cell source for meniscus repair. BMC Musculoskelet. Disord. 2015, 16, 14. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Vasicek, J.; Kovac, M.; Balazi, A.; Kulikova, B.; Tomkova, M.; Olexikova, L.; Curlej, J.; Bauer, M.; Schnabl, S.; Hilgarth, M.; et al. Combined approach for characterization and quality assessment of rabbit bone marrow-derived mesenchymal stem cells intended for gene banking. New Biotechnol. 2020, 54, 1–12. [Google Scholar] [CrossRef]
- Barraud, P.; Stott, S.; Mollgard, K.; Parmar, M.; Bjorklund, A. In vitro characterization of a human neural progenitor cell coexpressing SSEA4 and CD133. J. Neurosci. Res. 2007, 85, 250–259. [Google Scholar] [CrossRef]
- Vasicek, J.; Balazi, A.; Chrenek, P. The efficiency of immunomagnetic sorting of rabbit bone marrow cells for the establishment of mesenchymal stem cell culture. J. Microbiol. Biotechnol. Food Sci. 2018, 8, 890–892. [Google Scholar] [CrossRef]
- Futerman, A.H.; Hannun, Y.A. The complex life of simple sphingolipids. EMBO Rep. 2004, 5, 777–782. [Google Scholar] [CrossRef] [Green Version]
- Merrill, A.H. De novo sphingolipid biosynthesis: A necessary, but dangerous, pathway. J. Biol. Chem. 2002, 277, 25843–25846. [Google Scholar] [CrossRef] [Green Version]
- Shayman, J.A. Sphingolipids. Kidney Int. 2000, 58, 11–26. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sobiesiak, M.; Sivasubramaniyan, K.; Hermann, C.; Tan, C.; Orgel, M.; Treml, S.; Cerabona, F.; de Zwart, P.; Ochs, U.; Muller, C.A.; et al. The Mesenchymal Stem Cell Antigen MSCA-1 is Identical to Tissue Non-specific Alkaline Phosphatase. Stem Cells Dev. 2010, 19, 669–677. [Google Scholar] [CrossRef]
- Jasper, P.J.; Zhai, S.K.; Kalis, S.L.; Kingzette, M.; Knight, K.L. B lymphocyte development in rabbit: Progenitor B cells and waning of B lymphopoiesis. J. Immunol. 2003, 171, 6372–6380. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sivasubramaniyan, K.; Harichandan, A.; Schilbach, K.; Mack, A.F.; Bedke, J.; Stenzl, A.; Kanz, L.; Niederfellner, G.; Buhring, H.J. Expression of stage-specific embryonic antigen-4 (SSEA-4) defines spontaneous loss of epithelial phenotype in human solid tumor cells. Glycobiology 2015, 25, 902–917. [Google Scholar] [CrossRef] [Green Version]
- Shahaf, G.; Zisman-Rozen, S.; Benhamou, D.; Melamed, D.; Mehr, R. B Cell Development in the Bone Marrow Is Regulated by Homeostatic Feedback Exerted by Mature B Cells. Front. Immunol. 2016, 7, 13. [Google Scholar] [CrossRef] [Green Version]
- McElroy, P.J.; Willcox, N.; Catty, D. Early precursors of B lymphocytes I. Rabbit/mouse species differences in the physical properties and surface phenotype of pre-B cells, and in the maturation sequence of early B cells. Eur. J. Immunol. 1981, 11, 76–85. [Google Scholar] [CrossRef]
- Crane, M.A.; Kingzette, M.; Knight, K.L. Evidence for limited B-lymphopoiesis in adult rabbits. J. Exp. Med. 1996, 183, 2119–2127. [Google Scholar] [CrossRef] [Green Version]
- Mage, R.G.; Lanning, D.; Knight, K.L. B cell and antibody repertoire development in rabbits: The requirement of gut-associated lymphoid tissues. Dev. Comp. Immunol. 2006, 30, 137–153. [Google Scholar] [CrossRef] [PubMed]
- Kalis, S.L.; Zhai, S.K.; Yam, P.C.; Witte, P.L.; Knight, K.L. Suppression of B lymphopoiesis at a lymphoid progenitor stage in adult rabbits. Int. Immunol. 2007, 19, 801–811. [Google Scholar] [CrossRef] [Green Version]
- Kovac, M.; Vasicek, J.; Kulikova, B.; Bauer, M.; Curlej, J.; Balazi, A.; Chrenek, P. Different RNA and protein expression of surface markers in rabbit amniotic fluid-derived mesenchymal stem cells. Biotechnol. Prog. 2017, 33, 1601–1613. [Google Scholar] [CrossRef]
- Vasicek, J.; Shehata, M.; Schnabl, S.; Hilgarth, M.; Hubmann, R.; Jager, U.; Bauer, M.; Chrenek, P. Critical Assessment of the Efficiency of CD34 and CD133 Antibodies for Enrichment of Rabbit Hematopoietic Stem Cells. Biotechnol. Prog. 2018, 34, 1278–1289. [Google Scholar] [CrossRef] [PubMed]
- Kulikova, B.; Kovac, M.; Bauer, M.; Tomkova, M.; Olexikova, L.; Vasicek, J.; Balazi, A.; Makarevich, A.V.; Chrenek, P. Survivability of rabbit amniotic fluid-derived mesenchymal stem cells post slow-freezing or vitrification. Acta Histochem. 2019, 121, 491–499. [Google Scholar] [CrossRef]
- Ye, J.; Coulouris, G.; Zaretskaya, I.; Cutcutache, I.; Rozen, S.; Madden, T.L. Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinform. 2012, 13, 11. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pfaffl, M.W. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001, 29, 6. [Google Scholar] [CrossRef] [PubMed]
- Vašíček, J.; Baláži, A.; Bauer, M.; Svoradová, A.; Tirpáková, M.; Ondruška, Ľ.; Parkányi, V.; Makarevich, A.V.; Chrenek, P. Enrichment of Rabbit Primitive Hematopoietic Cells via MACS Depletion of CD45+ Bone Marrow Cells. Magnetochemistry 2021, 7, 11. [Google Scholar] [CrossRef]
- Vasicek, J.; Balazi, A.; Bauer, M.; Svoradova, A.; Tirpakova, M.; Tomka, M.; Chrenek, P. Molecular Profiling and Gene Banking of Rabbit EPCs Derived from Two Biological Sources. Genes 2021, 12, 366. [Google Scholar] [CrossRef] [PubMed]
- Sanchez-Rodriguez, A.; Arias-Alvarez, M.; Rebollar, P.G.; Bautista, J.M.; Lorenzo, P.L.; Garcia-Garcia, R.M. Gene expression and immunolocalization of low-affinity neurotrophin receptor (p75) in rabbit male reproductive tract during sexual maturation. Reprod. Domest. Anim. 2018, 53, 62–65. [Google Scholar] [CrossRef]
Sample | Con (SSEA-4+) | Neg (SSEA-4+) | Pos (SSEA-4+) | Pos (SSEA-4+CD45+) | Pos (SSEA-4+LCR+) |
---|---|---|---|---|---|
PBMCs | 58.6 ± 16.0 | 8.4 ± 2.4 ** | 90.1 ± 4.4 * | 89.5 ± 3.4 | 88.6 ± 3.9 |
BMMCs | 26.4 ± 9.1 | 5.2 ± 1.5 * | 84.8 ± 1.8 ** | 82.5 ± 3.4 | 80.7 ± 3.0 |
Marker | Host/Isotype | Clone | Conjugate | Company |
---|---|---|---|---|
CD4 | mouse IgG1 | RTH1A | purified 1 | WSU |
CD8 | mouse IgG2a | ISC27A | purified 1 | WSU |
CD14 | mouse IgG2a | TÜK4 | FITC | Dako Cytomation |
CD45 | mouse IgG1 | L12/201 | purified 1 | Bio-Rad |
CD45 | mouse IgG2a | ISC18A | purified 1 | WSU |
IgM | mouse IgG1 | NRBM | purified 1 | Bio-Rad |
pan T2 | mouse IgG1 | RTH21A | purified 1 | WSU |
SSEA-4 | mouse IgG3 | MC-813-70 | PE | eBioscience |
Gene | Product Size (bp) | Forward Primer | Reverse Primer | Primer Efficiency | Reference |
---|---|---|---|---|---|
ST3GAL2 (SSEA-4) | 126 | 5′-CTGGGAGAATAACCGGTACG-3′ | 5′-GCTCAGTTGCCTCGGTAGAC-3′ | 2.04 | [19] |
CD45 | 262 | 5′-TACTCTGCCTCCCGTTG-3′ | 5′-GCTGAGTGTCTGCGTGTC-3′ | 2.02 | [19] |
CD14 | 128 | 5′-TCTCTGTCCCCACAAGTTCC-3′ | 5′-GGCTGAGGTCTAGGTGATGG-3′ | 1.92 | NM_001082195.2 1 |
CD79α | 155 | 5′-CATCGGAAGTACGGAGCATT-3′ | 5′-TCCTTCCAGTCATCCCACTC-3′ | 2.06 | XM_008272740.2 1 |
IGHM (IgM) | 127 | 5′-GCCTGTACTTCACCCACAGC-3′ | 5′-GACTTGTCCACGGTCCTCTC-3′ | 2.04 | MN651044.1 1 |
CD3g | 104 | 5′-TCATTGCAGGACAAGATGGA-3′ | 5′-GTCATCTTCCCGATCCTTGA-3′ | 2.01 | XM_002722697.3 1 |
CD4 | 163 | 5′-CAGTCCTGGGTCAGCTTCTC-3′ | 5′-TATCAAGGGTCAGGCTCAGG-3′ | 2.06 | NM_001082313.2 1 |
CD8a | 133 | 5′-GTGGACTCTCCGCTCAACTC-3′ | 5′-GGCTGAAGTGTAGGCTGAGG-3′ | 2.07 | XM_017340260.1 1 |
CD8b | 113 | 5′-CAACCCGGTCTGTTCTCAGT-3′ | 5′-TGTCAGCTGAGTTCCCTTCC-3′ | 2.05 | XM_008254148.2 1 |
MHCI | 97 | 5′-AGTGGGAATTGTTGCTGGAG-3′ | 5′-TCCTTTCCCATCTGAGCTGT-3′ | 1.91 | [38] |
MHCII | 142 | 5′-CTGTGAGCACAGGAAGGTGA-3′ | 5′-GGGTGGCTAGATGTCTGGAA-3′ | 2.00 | [38] |
B2M | 118 | 5′-ATTCACGCCCAATGATAAGG-3′ | 5′-ATCCTCAGACCTCCATGCTG-3′ | 2.02 | [19] |
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Vašíček, J.; Baláži, A.; Bauer, M.; Chrenek, P. SSEA-4 Antigen Is Expressed on Rabbit Lymphocyte Subsets. Magnetochemistry 2021, 7, 94. https://0-doi-org.brum.beds.ac.uk/10.3390/magnetochemistry7070094
Vašíček J, Baláži A, Bauer M, Chrenek P. SSEA-4 Antigen Is Expressed on Rabbit Lymphocyte Subsets. Magnetochemistry. 2021; 7(7):94. https://0-doi-org.brum.beds.ac.uk/10.3390/magnetochemistry7070094
Chicago/Turabian StyleVašíček, Jaromír, Andrej Baláži, Miroslav Bauer, and Peter Chrenek. 2021. "SSEA-4 Antigen Is Expressed on Rabbit Lymphocyte Subsets" Magnetochemistry 7, no. 7: 94. https://0-doi-org.brum.beds.ac.uk/10.3390/magnetochemistry7070094