skip to main content
Idioma:

Isolation and culture of murine B1A cells

F G Thies Mário Mariano; A F Popi; Meeting of the Brazilian Society for Immunology (31. 2006 Búzios)

Abstracts São Paulo, SP: Brazilian Society for Immunology, 2006

São Paulo 2006

Item não circula. Consulte sua biblioteca.(Acessar)

  • Título:
    Isolation and culture of murine B1A cells
  • Autor: F G Thies
  • Mário Mariano; A F Popi; Meeting of the Brazilian Society for Immunology (31. 2006 Búzios)
  • Assuntos: IMUNOLOGIA
  • É parte de: Abstracts São Paulo, SP: Brazilian Society for Immunology, 2006
  • Notas: Disponível em CD-ROM
  • Descrição: Introduction and Objectives: B-1 cells differ from conventional B cells by their origin, anatomic localization, and surface markers expression. There are two kinds of B-1 cell: B-1b (CD23-CD19+CD11b+CD5-) and B-1a (CD23-CD19+CD11b+CD5+). It has been demonstrated in our laboratory that, in a culture of total peritoneal cells, B-1a cells are the non-adherent fraction, as opposed to B-1b cells, which adhere to the culture plate. B-1a cells are not able to adhere even when maintained for a long time in culture. As there is no information concerning the behavior of B-1a cells in culture the objective of this work is to standardize the isolation and culture of B-1a cells to further contribute to the origin and properties of these cells. Methods and Results: Total peritoneal cells from BALB/c mice were kept in culture for 40 minutes at 37ºC and 5% CO2. Adherent cells were discarded and non-adherent cells collected and re-cultured. After flow cytometric analysis using monoclonal antibodies conjugated with different fluorocromes, we noted two distinct populations in the culture: CD5+CD11b+CD19+CD23- (B-1a cells) and CD5+CD19-CD11b-CD23-. Both cell populations were kept in culture for 7 days. The CD19+ fraction increased until the third day in culture. Conversely, there was not a significant increase in CD19- cells. CD19+ cells were isolated using magnetic beads and stained with CFSE (2mM), showing that 40.6% of CD19+ cells were multiplying
    until the third day in culture. IgM production by these cells is under investigation. Besides, a T cell contamination was observed in the culture, with 3.5% of CD3+CD5+CD4+CD8- cells and 7.5% of CD3+CD5+CD4+CD8- cells. Conclusions: These data led us to conclude a) that B-1a cells are not adherent to the glass surface and proliferate up to the third day in culture b) that B-1 b cells were not found in these cultures and c) that these cultures are "contaminated" with an unknown cell type (CD5+CD19-CD11b-CD23-) and with a small proportion of T cells.
  • Editor: São Paulo
  • Data de criação/publicação: 2006
  • Formato: res. CI.067.
  • Idioma: Inglês
Links
Voltar para lista de resultados
Resultado  1

  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

Buscando em bases de dados remotas. Favor aguardar.