Abstract: Visible and near infrared (VIS-NIR) spectroscopy has been applied to estimate soil organic carbon (SOC) content with many modeling strategies and techniques, in which a crucial and challenging problem is to obtain accurate estimations using a limited number of samples with reference values (labeled samples). To solve such a challenging problem, this study, with Honghu City (Hubei Province, China) as a study area, aimed to apply semi-supervised regression (SSR) to estimate SOC contents from VIS-NIR spectroscopy. A total of 252 soil samples were collected in four field campaigns for laboratory-based SOC content determinations and spectral measurements. Semi-supervised regression with co-training based on least squares support vector machine regression (Co-LSSVMR) was applied for spectral estimations of SOC contents, and it was further compared with LSSVMR. Results showed that Co-LSSVMR could improve the estimations of SOC contents by exploiting samples without reference values (unlabeled samples) when the number of labeled samples was not excessively small and produce better estimations than LSSVMR. Therefore, SSR could reduce the number of labeled samples required in calibration given an accuracy threshold, and it holds advantages in SOC estimations from VIS-NIR spectroscopy with a limited number of labeled samples. Considering the increasing popularity of airborne platforms and sensors, SSR might be a promising modeling technique for SOC estimations from remotely sensed hyperspectral images.