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Critical assessment of the efficiency of CD34 and CD133 antibodies for enrichment of rabbit hematopoietic stem cells.
Vasícek, Jaromír; Shehata, Medhat; Schnabl, Susanne; Hilgarth, Martin; Hubmann, Rainer; Jäger, Ulrich; Bauer, Miroslav; Chrenek, Peter.
Afiliação
  • Vasícek J; NAFC-Research Institute for Animal Production in Nitra, Institute of Farm Animal Genetics and Reproduction, Luzianky, Slovak Republic, Hlohovecká 2, 951 41.
  • Shehata M; Research Centre AgroBioTech, Slovak University of Agriculture, Tr. A. Hlinku 2, 949 76 Nitra, Slovak Republic.
  • Schnabl S; Faculty of Biotechnology and Food Science, Department of Biochemistry and Biotechnology, Slovak University of Agriculture, Nitra, Tr A. Hlinku 2, 949 76, Slovak Republic.
  • Hilgarth M; Dept. of Internal Medicine I, Div. of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
  • Hubmann R; Dept. of Internal Medicine I, Div. of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
  • Jäger U; Dept. of Internal Medicine I, Div. of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
  • Bauer M; Dept. of Internal Medicine I, Div. of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
  • Chrenek P; Dept. of Internal Medicine I, Div. of Haematology and Haemostaseology, Comprehensive Cancer Centre Vienna, Drug and Target Screening Unit DTSU, Medical University of Vienna, Waehringer Guertel 18-20, Vienna, A-1090, Austria.
Biotechnol Prog ; 34(5): 1278-1289, 2018 09.
Article em En | MEDLINE | ID: mdl-29882300
ABSTRACT
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., 341278-1289, 2018.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Células-Tronco Hematopoéticas / Antígenos CD34 / Antígeno AC133 / Anticorpos Limite: Animals Idioma: En Ano de publicação: 2018 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Células-Tronco Hematopoéticas / Antígenos CD34 / Antígeno AC133 / Anticorpos Limite: Animals Idioma: En Ano de publicação: 2018 Tipo de documento: Article