diff --git a/ci/scripts/python_wheel_windows_test.bat b/ci/scripts/python_wheel_windows_test.bat
index cac3f18434b6c..1f0963eeaf7dd 100755
--- a/ci/scripts/python_wheel_windows_test.bat
+++ b/ci/scripts/python_wheel_windows_test.bat
@@ -64,6 +64,9 @@ set PYTHON_CMD=py -%PYTHON%
 %PYTHON_CMD% -c "import pyarrow.parquet" || exit /B 1
 %PYTHON_CMD% -c "import pyarrow.substrait" || exit /B 1
 
+@REM Validate wheel contents
+%PYTHON_CMD% C:\arrow\ci\scripts\validate_wheels_content.py --wheel C:\arrow\python\dist\*.whl || exit /B 1
+
 @rem Download IANA Timezone Database for ORC C++
 curl https://cygwin.osuosl.org/noarch/release/tzdata/tzdata-2024a-1.tar.xz --output tzdata.tar.xz || exit /B
 mkdir %USERPROFILE%\Downloads\test\tzdata