Assessment of different protocols for the isolation and purification of gut associated lymphoid cells from the gilthead seabream (Sparus aurata L.)

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Author: Salinas, I
Author: Esteban, M
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Methods Article

Assessment of different protocols for the isolation and purification of gut associated lymphoid cells from the gilthead seabream (Sparus aurata L.)

Irene Salinas¹, José Meseguer¹ and Maria Ángeles Esteban¹^*

¹ Fish Innate Immune System Group, Department of Cell Biology, Faculty of Biology, University of Murcia. 30100 Murcia. Spain. Phone: +34 968367665 Fax: +34 968363963 Email: aesteban@um.es.

* To whom correspondence should be addressed: Maria Ángeles Esteban, Fish Innate Immune System Group, Department of Cell Biology, Faculty of Biology, University of Murcia. 30100 Murcia. Spain. Phone: +34 968367665. Fax: +34 968363963. Email: aesteban@um.es

Biol. Proced. Online 2007;9:43-55. doi:10.1251/bpo132
Submitted: February 15, 2007; Accepted: March 30, 2007; Published: November 25, 2007.

Indexing terms: Sea Bream; Flow Cytometry; Respiratory Burst.

Figure 1 Enlarged

Fig. 1:

Flow cytometry SSC and FFC dot plots of seabream intestine cells showing three subpopulations (R1, R2 and R3). (a) Representative dot plot of a S2 suspension obtained after isolation of cells using both chemical (DTT, 10 min) and enzymatic (collagenase 0.15 mg/ml, 60 min) treatments. (b) Representative dot plot obtained after the same suspension in Figure 1a is passed through a nylon wool column (NW) for 60 min. (c) Representative dot plot of a total suspension obtained by mechanical stripping,

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