SHAPE CHANGES OF RED BLOOD CELLS DURING MECHANICAL STIRRING AND REPLACEMENT IN THE MEDIUM OF SULFATE TO CHLORIDE
V.V. Ramazanov, S.V. Rudenko
- Institute for Problems of Cryobiology and Cryomedicine of
the National Academy of Sciences of Ukraine, Kharkov, Ukraine
DOI: https://doi.org/10.15407/fz70.04.072
Abstract
At low pH, red blood cells (RBCs) are unable to maintain their
disc shape and become stomatocytes. At the same time, at pH
5.0 of a sulphate medium, under conditions of mechanical
mixing of the cell suspension, RBCs are partially retransformed
into discoid forms, whereas in a chloride medium (pH 5.0) this
transformation is observed to a high degree.This indicates
that the chloride-binding sites of RBC membranes may be an
additional link in the regulation of the shape of RBC under
conditions of mechanical action on the cell suspension. The
work investigated the change in the RBC shape at a normal
pH value (7,4) due to the replacement of Na2SO 4 in the
medium with NaCl and recording an increase in the intensity
of fluctuations in optical density (OD), as an indicator of an
increase in the level of discoid cells (normocytes) in a stirred
cell suspension. It has been 3 established that in a medium
containing Na2SO4 (110 mmol/l), RBCs are transformed into
small stomatocytes within ~20 s. With an increase in NaCl
concentration in the range of 15-105 mmol/l, an increase
in the level of discoid cells is noted. Increasing the NaCl
concentration from 105 to 150 mmol/l, on the contrary, causes
a decrease in the level of discoid cells. With the exception of
mechanical mixing in a medium with NaCl concentrations
of 30-90 mmol/l, RBCs are morphologically stomatocytes.
Whereas in a medium containing 150 mmol/l NaCl, RBC are
represented by disc-echinocytes and echinocytes. The results
obtained show that mechanical mixing of the cell suspension
promotes the retransformation of stomatocytic RBCs into
discoid forms. The weakening of the retransforming efficiency
of mixing with an increase in the NaCl concentration in the
medium from 105 to 150 mmol/l and the development of
echinocytosis at 150 mmol/l NaCl indicates the existence of
2 types of sites for chloride. Сhloride binding to type 1, at a
NaCl concentration in the medium of 15-105 mmol/l, leads to
the establishment of discoid forms of RBC. Chloride binding
to type 2 sites, at a NaCl concentration in the medium of
105-150 mmol/l, leads to the development of echinocytosis.
The retransforming property of mechanical stirring may be
associated with a change in the degree of chloride binding to
these types of sites.
Keywords:
red blood cells; RBC; chloride; sulfate; cell shape.
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