Isolation of rat lung mast cells for purposes of one-week cultivation using novel percoll variant percoll PLUS

Title:

Isolation of rat lung mast cells for purposes of one-week cultivation using novel percoll variant percoll PLUS

Creator:

Kubrycht, J., Hana Maxová, Otakar Nyč, and Luděk Vajner

Identifier:

https://cdk.lib.cas.cz/client/handle/uuid:48a0a79a-9f08-47e8-8b00-7961e495f418
uuid:48a0a79a-9f08-47e8-8b00-7961e495f418

Subject:

Fyziologie člověka a srovnávací fyziologie, hypoxie, hypoxia, lung mast cells, percoll, c-kit, progenitor, secretion, 14, and 612

Type:

article, články, model:article, and TEXT

Format:

print

Description:

Prolonged cultivation of separated rat lung mast cells (LMC) in vitro is necessary to better investigate a possible role of LMC in different stages of tissue remodeling induced by hypoxia. Rat lung mast cells (LMC) were sepa rated using a protocol including an improved proteolytic extracti on and two subsequent density gradient separations on Ficoll-P aque PLUS and a new generation of Percoll, i.e. Percoll PLUS. Instead of usual isotonic stock Percoll solution, an alternative “asymptotically isotonic” stock solution was more successful in our density separation of LMC on Percoll PLUS. Separated cells were cultivated for six days in media including stem cell factor, interleu kins IL-3 and IL-6, and one of two alternative mixtures of antibi otics. These cultivations were performed without any contaminatio n and with only rare changes in cell size and morphology. Model co-cultivation of two allogenic fractions of LMC often caused considerable rapid changes in cell morphology and size. In contrast to these observations no or rare morphological changes were found after cultivation under hypoxic conditions. In conclusions, we modified separation on Percoll PLUS to be widely used, altered LMC separation with respect to purposes of long-lasti ng cultivation and observed some model morphological changes of LMC., J. Kubrycht ... [et al.]., and Obsahuje bibliografii a bibliografické odkazy

Language:

English

Rights:

http://creativecommons.org/licenses/by-nc-sa/4.0/
policy:public

Source:

Physiological research | 2011 Volume:60 | Number:1

Harvested from:

CDK

Metadata only:

false