Critical assessment of the efficiency of CD34 and CD133 antibodies for enrichment of rabbit hematopoietic stem cells
| Autor: | Jaromír Vašíček, Peter Chrenek, Susanne Schnabl, Martin Hilgarth, Ulrich Jäger, Medhat Shehata, Rainer Hubmann, Miroslav Bauer |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
medicine.drug_class Antigens CD34 Monoclonal antibody Antibodies Flow cytometry 03 medical and health sciences 0302 clinical medicine medicine Animals AC133 Antigen biology medicine.diagnostic_test Chemistry Hematopoietic stem cell Cell sorting Flow Cytometry Hematopoietic Stem Cells Molecular biology 030104 developmental biology medicine.anatomical_structure Polyclonal antibodies 030220 oncology & carcinogenesis biology.protein Rabbits Antibody Stem cell Clone (B-cell biology) Biotechnology |
| Zdroj: | Biotechnology Progress. 34:1278-1289 |
| ISSN: | 1520-6033 8756-7938 |
| Popis: | Rabbits have many hereditary diseases common to humans and are therefore a valuable model for regenerative disease and hematopoietic stem cell (HSC) therapies. Currently, there is no substantial data on the isolation and/or enrichment of rabbit HSCs. This study was initiated to evaluate the efficiency of the commercially available anti-CD34 and anti-CD133 antibodies for the detection and potential enrichment of rabbit HSCs from peripheral blood. PBMCs from rabbit and human blood were labelled with different clones of anti-human CD34 monoclonal antibodies (AC136, 581, and 8G12) and rabbit polyclonal CD34 antibody (pCD34) and anti-human CD133 monoclonal antibodies (AC133 and 293C3). Flow cytometry showed a higher percentage of rabbit CD34+ cells labelled by AC136 in comparison to the clone 581 and pCD34 (P < 0.01). A higher percentage of rabbit CD133+ cells were also detected by 293C3 compared to the AC133 clone (P < 0.01). Therefore, AC136 clone was used for the indirect immunomagnetic enrichment of rabbit CD34+ cells using magnetic-activated cell sorting (MACS). The enrichment of the rabbit CD34+ cells after sorting was low in comparison to human samples (2.4% vs. 39.6%). PCR analyses confirmed the efficient enrichment of human CD34+ cells and the low expression of CD34 mRNA in rabbit positive fraction. In conclusion, the tested antibodies might be suitable for detection, but not for sorting the rabbit CD34+ HSCs and new specific anti-rabbit CD34 antibodies are needed for efficient enrichment of rabbit HSCs. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018 © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1278-1289, 2018. |
| Databáze: | OpenAIRE |
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