Separation of Peripheral Leukocytes by Ficoll Density Gradient Centrifugation

Tripodi, D; Lyons, Susan F.; Davies, Donald M.

doi:10.1097/00007890-197105000-00010

Cited by 27 publications

(15 citation statements)

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“…[15--17, 41, 48--51]. Thus, preparation of lymphocytes [59], density of cells in the culture suspension [15,17,41], and the culture medium itself, including serum content and the duration of cultivation in the presence of PHA are reasonably well established. However, stimulation by PHA first requires the use of a single charge of mitogen with dilutions prepared in one operation, and second the regular elaboration of a dose response curve, in order to register differences to stimulation between the examined groups.…”

Section: Discussionmentioning

confidence: 99%

Lymphocyte proliferation to Phytohemagglutinin (PHA) in hepatitis B antigen-positive and -negative hepatitis

et al. 1979

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Lymphocytes from patients with HBs-Ag-positive and -negative acute, chronic-persistent, and chronic-active hepatitis, from healthy controls and from patients with alcoholic liver cirrhosis were tested under standardized conditions. These included use of a single charge of Phytohemagglutinin (PHA-P) dissolved and diluted in one operation, of a single pool of homologous serum of the major blood group AB found free of HBs-Ag and cytotixic factor, and elaboration of PHA dose response curves in the presence of autologous and homologous serum in each case examined. During the early phase of acute virus hepatitis B and non-B, and in HBs-Ag-positive chronic persistent and active hepatitis, hyperresponsiveness of lymphocytes to PHA was observed independently of the source of the serum present in the culture. Lymphocyte responsiveness returned to normal in the later phase of acute hepatitis and depressed in alcoholic liver cirrhosis and in cases of HBs-Ag-positive chronic active hepatitis in which cirrhosis had developed. Although the cause of these alterations in lymphocyte responsiveness is not completely understood, the central role of a primary change of the lymphocytes themselves affecting their ability to react to PHA seems probable.

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Section: Discussionmentioning

confidence: 99%

Lymphocyte proliferation to Phytohemagglutinin (PHA) in hepatitis B antigen-positive and -negative hepatitis

et al. 1979

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“…Samples were diluted 1:2 with RPMI medium 1640 supplemented with 10% FCS and penicillin/streptomycin (50 U/mL and 50 g/mL). Lymphocytes were separated using gradient centrifugation by layering a total of 4 mL of diluted blood onto 4 mL of Histopaque 1077 (Sigma-Aldrich, Munich, Germany) and centrifuged at 1200g for 15 min [29]. This separation yields approximately 80% lymphocytes, 15% monocytes and 5% granulocytes, as described before [28].…”

Section: Conventional Coronary Angiographymentioning

confidence: 99%

DNA double-strand breaks as potential indicators for the biological effects of ionising radiation exposure from cardiac CT and conventional coronary angiography: a randomised, controlled study

et al. 2012

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“…Lymphocyte separation was performed by using Ficoll-Hypaque densitygradient centrifugation with lymphocyte separation medium 1077 (PAA Laboratories) (22). Blood samples (5 mL diluted) were carefully layered onto lymphocyte separation medium and centrifuged at 1200g for 20 minutes at room temperature.…”

Section: Sample Processing and Lymphocyte Separationmentioning

confidence: 99%