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Bone Marrow Mononuclear Cell Extraction. Does the Operator Performance Affect Processing Efficacy？

Journal of Pharmacy and Pharmacology 2016年第5期4卷 192-198页

作者： Jakobsons Eriks Cakstina Inese Erglis Kristaps Erglis Martins pirtniece liene Elina Krumina NarbuteInga Briede Ieva Jegere Sanda Smolova Rita Muiznieks Indrikis Erglis Andrejs Latvian Centre of Cardiology Pauls Stradins Clinical University Hospital Riga L V-1002 Latvia Cell Transplantation Centre Pauls Stradins Clinical University Hospita l Riga LV-1002 Latvia Research Institute of Cardiology University of Latvia Riga LV-1586 Latvia Department of Biology Division of Microbiology and Bioteehnology University of Latvia Riga LV-1586 Latvia

Different cell populations from bone marrow are used in various clinical trials for cardiac diseases during last decade. Four clinical studies are on going in our institution and enrol patients with cardiac diseases, coronary disease and type 2 diabetes, patients with osteoarthritis. Density gradient is used to separate bone marrow mononuclear cells. Cell processing looses are significant. To find out critical control points we analysed processing process and differences in cell yields between operators performing cell extraction. Bone marrow mononuclear cells were isolated using Ficoll density gradient centrifugation. Cells were counted using flow cytometry for mononuclear cell total counts, CD34＋ population count and viability analysis. The patients who underwent bone marrow aspiration followed by cell isolation received cell suspension for transplantation. Two cells processing for separate patients were performed at once. Same standard operation procedures were applied. Processing looses between operators performing cell extraction were analysed. Bone marrow samples from eight patients were processed. Mononuclear cells were extracted. Operator performances were compared. Operator A average bone marrow mononuclear cell yield in starting material was 9,97 ± 9,98 %, CD34＋ population yield 75,46±79,67%. Operator B average bone marrow mononuclear cell yield in starting material was 24,68 ± 14,8 %, CD34＋ population yield 70,42 ±44.84%. Operator A average cell viability in starting material was 45,24 ± 9,55%, after cell processing 42,96 ± 23,66 %. Operator B average cell viability in starting material was 49,85 ± 5,48%, after cell processing 69,52 ± 6,65 %.

关键词： Bone marrow mononuclear cells cell yield cell extraction cell count

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