LCAT, ApoD and ApoA1 were immunolocalized to keratocytes in the cornea. RNA sequencing analysis of cultured keratocytes revealed the presence of both LCAT and ApoD mRNA, which was accompanied by their respective proteins detected by keratocyte immunolabeling and Western blot analysis of keratocyte lysates. In contrast, ApoA1 mRNA was absent and cultured keratocytes, in contrast to in vivo keratocytes, did not show immunostaining for ApoA1. Thus, keratocytes can synthesize both LCAT and ApoD, but not ApoA1. However, in vivo, keratocytes presumably accumulate and store ApoA1 from the tissue fluid that perfuses the cornea. In situ hybridization analysis of LCAT expression in the cornea showed that cornea epithelium and endothelium, the two cell types surrounding the stroma anteriorly and posteriorly, respectively, synthesize LCAT to a much greater extent than keratocytes in the cornea. Our results suggest that LCAT locally produced within the cornea is important for cholesterol mobilization from this connective tissue. Thus, genetic loss of LCAT expression by cells of the cornea may greatly affect cholesterol removal from this tissue.