Respiratory Activity of Intact, Isolated Parenchymal Cells from Rat Liver

Howard, Roger B.; Pesch, Leroy A.

doi:10.1016/s0021-9258(18)93383-1

Cited by 189 publications

(12 citation statements)

References 13 publications

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“…The use of 100% oxygen is preferred to the mixture of 95% oxygen and 5% CO2 previously described (16,17) . The reason for this is that it is easier to maintain a pH close to 7 .4.…”

Section: Preparation Of Cell Suspensionsmentioning

confidence: 99%

“…We have already reported some modifications of our original technique . These resulted in the production of suspensions containing a high percentage of intact and viable cells so that calcium ions stimulated oxygen uptake (17) instead of depressing it as had occurred with other liver cell preparations (2,19) .…”

Section: Introductionmentioning

confidence: 99%

“…We have noted that our previously described procedure (16,17) in the hands of some workers has resulted in the preparation of damaged cells (4,10,15,20) . Since there are many critical factors involved in obtaining intact, isolated liver parenchymal cells, we are describing them in detail together with a complete account of our latest modified procedure .…”

Section: Introductionmentioning

confidence: 99%

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The Fine Structure, Potassium Content, and Respiratory Activity of Isolated Rat Liver Parenchymal Cells Prepared by Improved Enzymatic Techniques

Howard

Lee

Pesch

1973

The Journal of Cell Biology

103

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Further modifications of the enzymatic technique for the preparation of isolated, intact, parenchymal cells from rat liver as previously described by this laboratory are presented together with a detailed account of several critical factors involved during the procedure . In addition, the fine structure of the cells as revealed by electron microscopy and the characteristics of their respiratory activity in different media and with several added substrates are described . It is shown that cells obtained by adding calcium during the preparative procedure retain approximately 34% more potassium than cells prepared solely in a calcium-free medium . The former cells also demonstrate a higher respiratory activity, which is not due to uncoupling of respiration . Electron microscopy reveals that the cells have an intact plasma membrane and well-preserved intracellular organelles . Glycogen particles are observed in all cells and are particularly abundant when either 20 mM pyruvate is added during the preparation or Eagle's Minimum Essential Medium is employed .

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“…The use of 100% oxygen is preferred to the mixture of 95% oxygen and 5% CO2 previously described (16,17) . The reason for this is that it is easier to maintain a pH close to 7 .4.…”

Section: Preparation Of Cell Suspensionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

The Fine Structure, Potassium Content, and Respiratory Activity of Isolated Rat Liver Parenchymal Cells Prepared by Improved Enzymatic Techniques

Howard

Lee

Pesch

1973

The Journal of Cell Biology

103

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“…Cultures were split (1:3 v/v) when reaching 80% confluence; culture medium was replenished every 3 days. The trypan blue exclusion assay (Howard andPesch, 1968, Hauton andSmith, 2004) was used to determine cell viability and only cell suspensions with at least 95% viability were used for experimentation.…”

Section: A C C E P T E D Accepted Manuscriptmentioning

confidence: 99%

In vitro investigations on the effects of semi-synthetic, sulphated carbohydrates on the immune status of cultured common carp (Cyprinus carpio) leucocytes

Kareem1

Yates

Skidmore

et al. 2018

Fish & Shellfish Immunology

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The rapid emergence of drug resistance, unfavourable immunosuppression and mounting evidence to suggest the deleterious accumulation of drug breakdown residues within animal tissues has driven a strong desire to move away from these current methods of disease control. Some natural products such as β-glucan, which are extracted from, for example, plants and fungi, are able to modulate the immune system and increase protection against diseases. However, these products are heterogeneous and their effects can be variable thus limiting their applicability and reliability. Carbohydrates were modified via chemical sulphation and these semi-synthetic, sulphated carbohydrates analysed for their immunological activity utilising carp pronephric cells and a carp leucocyte cell line (CLC). A sulphated β(1,4)-glucan, methyl hydroxyethyl cellulose sulphate (MHCS), demonstrated a stimulatory effect on fish immune cells. MHCS induced a range of bioactive effects in carp leucocyte cells whilst not affecting cell viability when cells were exposed for 24 h at concentrations of 1-150 μgml. MHCS stimulated the innate immune system where a significant increase in respiratory burst activity was observed at concentrations 25-250 μgml in comparison to control (sterile water), cellulose ether, MacroGard and zymosan. Also, under in mock bacterial and viral infection conditions i.e. Lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (Poly(I:C)), MHCS enhanced the immune responses of pronephric cells by stimulating the respiratory burst activity at concentrations 50 and 150 μgml. MHCS also enhanced the expression of cytokines including interleukin 1 beta (IL1β), tumor necrosis factor alpha 1 and 2 (TNFα 1,2), interferons alpha 2 (IFN α2) and inducible nitric oxide synthase (iNOS) in carp pronephric cells. It is proposed that this new semi-synthetic carbohydrate is a potential candidate for the development of a new generation of immunostimulants and adjuvants for use in vaccination strategies in aquaculture.

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“…The methodology of liver cell cultures has been improved over the last years and currently, a variety of isolation, culture, and cryopreservation models have been established and evaluated (99). Shortly after the rst description of hepatocyte isolation by collagenase-and hyaluronidase-dissociatio n (61,62), new improvements were made by perfusing the livers in situ with the dissociating agents (12). Later, the protocol of the 2-step collagenase technique for rat hepatocytes has been established (84,85), which was later modi ed (98) and works well for short-term cultures of hepatocytes of a variety of species including rat, monkey, pig, dog, rabbit, and human.…”

Section: Isolated Liver Cell Modelsmentioning

confidence: 99%

In Vitro Models to Study Hepatotoxicity

Groneberg¹,

Große-Siestrup

Fischer³

2002

Toxicol Pathol

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Drug discovery and developmen t consists of a series of processes starting with the demonstration of pharmacologica l effects in experimental cell and animal models and ending with drug safety and ef cacy studies in patients. A main limitation is often the unacceptabl e level of toxicity with the liver as the primary target organ. Therefore, approache s to study hepatic toxicity in the early phase of drug discovery represent an important step towards rational drug development . A variety of in vitro liver models have been developed in the past years. Next to their use in drug development , they can also be applied to study environmenta l toxins and their hepatotoxicity. The 3 main approache s are ex vivo isolated and perfused organ models, precision-cut liver slices and cell culture models. Although the advantage of whole organ perfusions is based on the assessment of physiologic parameters such as bile production and morphologi c parameters such as tissue histology, cell culture models can be ef ciently used to assess cellular metabolism, cytotoxicity and genotoxicit y. The advantage of precision-cut liver slices is based on the juxtaposition of cellular assays and tissue morpholog y. None of these models can be compared as they all focus on different elds of hepatoxicolog y. For the future, the ideal setup for testing the hepatic toxicity of a new compoun d could of primary studies in cell or slice cultures to assess cellular effects and secondary studies using ex vivo perfused organs to examine gross organ function parameters and histology.

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Respiratory Activity of Intact, Isolated Parenchymal Cells from Rat Liver

Cited by 189 publications

References 13 publications

The Fine Structure, Potassium Content, and Respiratory Activity of Isolated Rat Liver Parenchymal Cells Prepared by Improved Enzymatic Techniques

The Fine Structure, Potassium Content, and Respiratory Activity of Isolated Rat Liver Parenchymal Cells Prepared by Improved Enzymatic Techniques

In vitro investigations on the effects of semi-synthetic, sulphated carbohydrates on the immune status of cultured common carp (Cyprinus carpio) leucocytes

In Vitro Models to Study Hepatotoxicity

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