A new method for the preparation of liver cells in suspension

S T JACOB & P M BHARGAVA

Published in: Experimental Cell Research, 1962, 27, 453-467.

Highlights:

1. A new method, in which a rubber pestle is used to disperse the tissue, has been described for preparation of rat liver cell suspensions.

2. An evaluation of the advantages and disadvantages of methods used in the past for preparing liver cell suspensions is presented.

3. The advantages of the new method, which is believed to be superior to all others previously described, are stated. The method yields a suspension which is free of blood cells, other cellular contaminants and cell debris, and in which the hepatic cord cells of the liver tissue are recovered in an almost quantitative yield. The method does not employ an enzymatic or a drastic physical treatment of the tissue at any stage, and takes only 15 min from the time the animal is sacrificed.

Abstract:

While a great deal of work has been done on the metabolism of tissue slices, relatively little is known about the metabolic behaviour of tissue cells in suspension, when they are not organised in patterns which are tissue specific. Information on the physical, chemical, biochemical and biological properties of tissue cells in suspension will be useful in many ways. For example, one approach towards understanding the biochemical and biophysical basis of intercellular organisation in tissues, in which we have been interested, would be to compare the metabolic behaviour and physical properties of tissue cells in suspension with those of the same cells when they are organised in a tissue, and to study the changes in the properties of individual cells during the various stages of reorganisation to give tissue-like structures. Such reorganisation of dissociated cells has been effectively achieved with several types of tissues, particularly embryonic, from a variety of animals [16, 38, 4 7, 48].

We have been interested in carrying out metabolic studies on tissue cells in suspension for another reason also. Some recent observations suggest a variation in some of the metabolic properties of individual mammalian cells which occur as free, single cells, as their concentration in cell suspension is varied. Thus, the rate of incorporation of amino acids into spermatozoa and reticulocyte proteins has been shown to vary inversely with the concentration of these cells in a suspension [ 10 ]. A similar concentration effect has been observed in regard to respiration of spermatozoa [11, 12]. This effect is unlikely .to be due to a nutrient being a limiting factor in these investigations, and it is possible that it may be caused by intercellular reactions, which, in solid tissues, may be related to growth and organisation. It will, therefore, be of some interest to study the variation, if any, in the metabolic properties of cells obtained in suspension from solid tissues, with their concentration in the suspension.

Suspensions of tissue cells also form excellent material for immunological studies seeking to elucidate the comparative role of various tissue factors in immunological response [48]. Such suspensions can, in addition, be used for cytological studies to yield information which cannot be easily or reliably obtained by studies on tissue slices [23].

For studies such as above, it is necessary that the tissue cells be obtained in suspension in biochemically, physically and structurally undamaged condition, except for such changes as would be incumbent to disorganisation. Moreover, the method used should give a good recovery of the tissue cells in the final, purified preparation, since a method which gives a poor recovery due to a breakage of the majority of the cells in unlikely to leave the recovered cells undamaged, although such damage may not be visually manifest. In addition, the suspensions must be free from blood cells and cell debris. In this paper, we report a method for the preparation of liver cell suspensions, which satisfies the above requirements. The relative merits and demerits of the methods previously described for the preparation of liver cell suspensions have also been discussed.

HOW TO CITE

A new method for the preparation of liver cells in suspension. S T JACOB & P M BHARGAVA. Experimental Cell Research, 1962, 27, 453-467.

READ THIS ARTICLE ONLINE OR ↓ DOWNLOAD A COPY

Download the full Paper from PMB memoir

↪ Link to Journal
➽ Full Text Online ↪ Download PDF

Having Trouble Downloading? ↪ Watch here how to download this article  ❙❙ ❚❚ ►

Tags: ,

Leave a Reply

Your email address will not be published. Required fields are marked *