Simple method for culture of peripheral blood lymphocytes of Testudinidae
| Autor: | Maria Tercília Vilela de Azeredo-Oliveira, Claudia Regina Bonini-Domingos, Maria Isabel Afonso da Silva, Larissa Paola Rodrigues Venancio, J.R. Santos, V.A.G. Moscheta, Carlos Eduardo Saranz Zago, Tiago Lucena da Silva, Luiz Dino Vizotto, A.V.B. Lima |
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| Přispěvatelé: | Universidade Estadual Paulista (Unesp) |
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Male
Lymphocyte Cell Culture Techniques Giemsa stain chemistry.chemical_compound Colchicine animal genetics chromosome Lymphocytes Genetics drug effect Karyotype General Medicine Turtles medicine.anatomical_structure female Female Brazil Mitotic index culture medium mitogenic agent phytohemagglutinin Biology lymphocyte chemistry Azure Stains colchicine Chromosomes Incubation period Fixatives mitosis index medicine Mitotic Index Animals Humans human Phytohemagglutinins Molecular Biology Mitosis turtle Molecular biology chromosome banding pattern culture technique Chromosome Banding Culture Media fixative Cell culture cytology Mitogens |
| Zdroj: | Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP |
| Popis: | Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:26:16Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:31:26Z : No. of bitstreams: 1 2-s2.0-84861829753.pdf: 510287 bytes, checksum: 59ee530bfb8c60f054141d21e88e74b6 (MD5) Made available in DSpace on 2014-05-27T11:26:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-12-01 We developed and optimized a simple, efficient and inexpensive method for in vitro culture of peripheral blood lymphocytes from the Brazilian tortoise Chelonoidis carbonaria (Testudinidae), testing various parameters, including culture medium, mitogen concentration, mitotic index, culture volume, incubation time, and mitotic arrest. Peripheral blood samples were obtained from the costal vein of four couples. The conditions that gave a good mitotic index were lymphocytes cultured at 37°C in minimum essential medium (7.5 mL), with phytohemagglutinin as a mitogen (0.375 mL), plus streptomycin/penicillin (0.1 mL), and an incubation period of 72 h. Mitotic arrest was induced by 2-h exposure to colchicine (0.1 mL), 70 h after establishing the culture. After mitotic arrest, the cells were hypotonized with 0.075 M KCl for 2 h and fixed with methanol/acetic acid (3:1). The non-banded mitotic chromosomes were visualized by Giemsa staining. The diploid chromosome number of C. carbonaria was found to be 52 in females and males, and sex chromosomes were not observed. We were able to culture peripheral blood lymphocytes of a Brazilian tortoise in vitro, for the preparation of mitotic chromosomes. |
| Databáze: | OpenAIRE |
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