| Autor: |
Silva TL; Departamento de Biologia, Centro de Estudos de Quelônios, Instituto de Biociências, Letras e Ciências Exatas, Universidade Estadual Paulista 'Júlio de Mesquita Filho', São José do Rio Preto, SP, Brasil. lucenabio@hotmail.com, Silva MI, Venancio LP, Zago CE, Moscheta VA, Lima AV, Vizotto LD, Santos JR, Bonini-Domingos CR, Azeredo-Oliveira MT |
| Jazyk: |
angličtina |
| Zdroj: |
Genetics and molecular research : GMR [Genet Mol Res] 2011 Dec 06; Vol. 10 (4), pp. 3020-5. Date of Electronic Publication: 2011 Dec 06. |
| DOI: |
10.4238/2011.December.6.2 |
| Abstrakt: |
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: |
MEDLINE |
| Externí odkaz: |
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