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Updated: Apr 17, 2026

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells

A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells

Published on: April 11, 2014

Ascorbic acid transport in brain microvascular pericytes.

William H Parker¹, Zhi-Chao Qu¹, James M May¹

¹Department of Medicine, Vanderbilt University School of Medicine, Nashville, TN 37232-0475, USA.

Biochemical and Biophysical Research Communications

|February 4, 2015

Summary

This summary is machine-generated.

Vascular pericytes utilize the Sodium-dependent Vitamin C Transporter 2 (SVCT2) to import vitamin C (ascorbic acid). This transporter mechanism is crucial for pericyte protection under diabetic conditions.

Keywords:

Ascorbic acid Pericytes SVCT2 Transport kinetics

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Area of Science:

Biochemistry
Cell Biology
Physiology

Background:

Intracellular vitamin C (ascorbic acid) protects vascular pericytes from apoptosis under simulated diabetic conditions.
The precise mechanism of ascorbate transport into pericytes remained unclear.

Purpose of the Study:

To elucidate the mechanism of ascorbate transport into vascular pericytes.
To identify the specific transporter responsible for intracellular vitamin C uptake in pericytes.

Main Methods:

Measuring intracellular ascorbate uptake kinetics in pericytes.
Utilizing transporter inhibitors (SVCT2 and GLUT inhibitors).
Western blot analysis and immunocytochemistry to confirm transporter presence and localization.

Main Results:

Pericytes exhibit linear ascorbate uptake with a Km of 21 μM, characteristic of Sodium-dependent Vitamin C Transporter 2 (SVCT2).
SVCT2 inhibition blocked ascorbate uptake, while GLUT inhibition affected dehydroascorbate uptake, confirming transporter specificity.
Western blot and immunocytochemistry confirmed SVCT2 expression on the pericyte plasma membrane and intracellularly.

Conclusions:

Sodium-dependent Vitamin C Transporter 2 (SVCT2) is the primary transporter for ascorbic acid in vascular pericytes.
Pericytes can concentrate intracellular ascorbate via an energy- and sodium-dependent process mediated by SVCT2.
These findings clarify ascorbate transport mechanisms in pericytes, relevant to diabetic vascular complications.