Google has quietly reduced the minimum audience size to just 100 users for all segment types across Search, Display, and YouTube campaigns. This update standardizes the eligibility threshold for Customer Lists, Remarketing Lists, and other audience segments, making it easier for smaller advertisers to activate audience-based targeting.
Additionally, the minimum size required for a segment to appear in Audience Insights has also been lowered from 1,000 to 100 users.

These changes were confirmed through the latest update to Google’s Audience Segments help documentation, which now includes added policy details and troubleshooting guidance.
This update was spotted by Dario Zannoni.