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The report from the II round of the quality control workshop for HLA typing

100%

Bogunia-Kubik K. , Laba A. , Lange A.

Postępy Higieny i Medycyny Doświadczalnej

2000

tom 54

nr 6

759-766

In november 1999 we started the quality control exercise for Polish institutions involved in HLA typing. Five out of eleven invited institutions responded favourably and took part in the first run of the workshop. In the second trial, presently reported, number of labs increased to eight. Each centre received 4 blood samples for serological typing of HLA class I and 5 samples for DNA HLA class II typing. As in the provious workshop, HLA class II typing should be performed as least for DRB1 alleles at the low resolution level. The paper presents the results of the second run of standardisation.

Characterization of human hepatocytes isolated from non-transplantable livers

88%

Laba A. , Ostrowska A. , Patrzalek D. , Paradowski L. , Lange A.

Archivum Immunologiae et Therapiae Experimentalis

2005

tom 53

nr 5

442-453

The successful use of hepatocytes depends on a reliable demonstration of the functional and morphological integrity of isolated cells. Herein we investigated whether the isolation and cryopreservation of primary human hepatocytes can compromise cell viability and liver-specific characteristics. Hepatocytes were isolated from encapsulated human liver segments by a modified 2-step perfusion technique. Isolated cells were Percoll-purified, cryopreserved, and stored in liquid nitrogen for 1?12 months. For rapid assessment of fresh and cryopreserve/thawed hepatocyte yield and viability, the cells were stained with trypan blue or labeled with fluorochromes. For immunocytochemical analysis, the cells were labeled with monoclonal antibodies for the presence of the following antigens and chemokines: CD3, CD45Ro, CD45Ra, CD34, CD68, CD90, CD95, CD20, HLA-DR, Ki67, PCNA, Bcl-2, p53, CXCR3, CXCR4, and SDF-1. The cells were tested for several specific functions, such as ureagenesis, energy status, MTT activity, lactate dehydrogenase leakage, and total CYP450 content. Assessment of both freshly isolated (Percoll-purified) and cryopreserved/thawed hepatocytes revealed a low constitutive level of contamination by non-parenchymal cells compared with crude (unpurified) preparations and tissue sections. All viable hepatocytes showed intact morphology and retained CYP450 protein, energy status, and urea synthesis. Modifications in hepatocyte preparations, such as depletion of dead, damaged, and non-parenchymal cells, improves cell purity, which can be adapted to further evaluation of hepatocyte immunogenicity. These data illustrate the importance and feasibility of human hepatocyte banking.

Human melanoma gene therapy: from animal studies to clinical trials

51%

Mackiewicz A. , Kapcinska M. , Laciak M. , Malicki J. , Murawa P. , Nowak , Sibilska E. , Wiznerowicz M. , Breborowicz D. , Lange A. , Hawley R. G.

Biotechnologia

1996

nr 4

42-54

Based on preclinical studies clinical protocol for immuno-gene therapy of human melanoma was designed in our Department.In January 1995, phase I clinical trial was initiated. Until now 5 patients with IV clinical degree of melanoma received geentic vaccine acco9rding to the following schedule: 4 injections in two weeks intervals, 3 injections in one month intervals and 3 injections in two month intervals.During therapy no toxic effects were observed.Induction of specific and non-specific anti-melanoma response was observed.Currently the trial enters phase II. Optimization of doses and immunization schedule as well as verification of patients eliglibility will be carried out.Moreover, clinical effcts of applied therapy will be monitored.