Isolation and culture of murine B1A cells (2006)
- Authors:
- Autor USP: MARIANO, MARIO - ICB
- Unidade: ICB
- Assunto: IMUNOLOGIA
- Language: Inglês
- Abstract: 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 multiplyinguntil 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.
- Imprenta:
- Source:
- Título: Abstracts
- Conference titles: Meeting of the Brazilian Society for Immunology
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ABNT
THIES, F G e MARIANO, Mário e POPI, A F. Isolation and culture of murine B1A cells. 2006, Anais.. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo, 2006. . Acesso em: 10 mar. 2026. -
APA
Thies, F. G., Mariano, M., & Popi, A. F. (2006). Isolation and culture of murine B1A cells. In Abstracts. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo. -
NLM
Thies FG, Mariano M, Popi AF. Isolation and culture of murine B1A cells. Abstracts. 2006 ;[citado 2026 mar. 10 ] -
Vancouver
Thies FG, Mariano M, Popi AF. Isolation and culture of murine B1A cells. Abstracts. 2006 ;[citado 2026 mar. 10 ] - Fagocitose de neutrofilos e seus granulos por macrofagos inflamatorios de camundongos das linhagens balb / c e c57bl/6
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