A single partitioning step in aqueous polymer two-phase systems reduces hypotonized rat erythrocyte heterogeneity.
Rat carrier erythrocytes prepared by hypotonic dialysis (80 mOsm/kg) are a heterogeneous cell population that can be fractionated into two well-defined cell subpopulations by a single partition step, in charge-sensitive dextran-poly(ethylene glycol) aqueous two-phase systems. One subpopulation (65% of total cells) has a decreased cell surface charge and is partitioned at the interface in a single step and then fractionated by counter-current distribution as a low-G subpopulation. The other subpopulation (35% of total cells) has charge surface properties more like those of the untreated control rat erythrocytes. These last cells are partitioned in the top phase in a single step and then fractionated by counter-current distribution as a high-G subpopulation. Partitioning is more effective in reducing cell heterogeneity in hypotonized rat erythrocyte populations than is density separation in Ficoll-paque which only separates a small less dense cell subpopulation (5% of total cells), with the most fragile cells, from a larger and more dense cell subpopulation (95% of total cells), with a mixture of fragile and normal cells. This simple cell separation procedure quickly reduces carrier erythrocyte heterogeneity in a single partitioning step so it can be used to prepare cells for in vivo studies.
Pérez
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MT
Departamento de Bioquímica y Biología Molecular, Universidad de Alcalá, Madrid, Spain.
García-Pérez
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AI
Lucas
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Sancho
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D000818:Animals; D002469:Cell Separation / Q000379:methods*; D003377:Countercurrent Distribution; D003911:Dextrans / Q000737:chemistry; D003956:Dialysis; D004912:Erythrocytes / Q000737:chemistry*; D006454:Hemoglobins / Q000032:analysis; D007038:Hypotonic Solutions; D011092:Polyethylene Glycols / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D051381:Rats; D012997:Solvents / Q000737:chemistry; D013499:Surface Properties
D003911:Dextrans; D006454:Hemoglobins; D007038:Hypotonic Solutions; D011108:Polymers; D012997:Solvents; D011092:Polyethylene Glycols