Effect Of Human Corneal Stromal Extract On Keratocytes From Smile-Derived Lenticules.
Published 2024 - 42nd Congress of the ESCRS
Reference: FP19.05 | Type: Free paper | DOI: 10.82333/q9e3-jz75
Authors: Yuexi Chen* 1 , Zheng Wang 2 , Jiansu Chen 3
1Refractive,Guangzhou Aier Eye Hospital,Guangzhou,China;Refractive,Aier Eye Institute,Changsha,China, 2Refractive,Guangzhou Aier Eye Hospital,Guangzhou,China, 3Refractive,Aier Eye Institute,Changsha,China
Purpose
To develop an effective approach for the proliferation of human corneal stromal cells (CSCs, also called keratocytes) derived from SMILE lenticules with preferable cell viability and physiological properties. Then, further, prove that these cells can be good seed cells for tissue engineering corneal construction and cell therapy.
Setting
The corneal stromal lenses produced during SMILE surgery are often discarded, while these lenses can serve as an effective source for obtaining human corneal stromal cells. Based on this, this study aims to investigate a method for in vitro cultivation of corneal stromal cells, which can maintain a quiet phenotype of cells and promote cell proliferation, thus meeting the needs of future corneal disease cell therapy and tissue engineering corneal construction.
Methods
We collected a large number of fresh SMILE lenticules and prepared soluble human corneal stromal extract (hCSE) with low-serum RIFA medium primarily including ROCK inhibitor Y27632, insulin-transferrin-selenium, fibroblast growth factor 2, L-ascorbate 2-phosphate and 0.5% fetal bovine serum (FBS) to culture primary CSCs obtained from SMILE-derived lenticules. The maintenance of the phenotype and physiological properties of keratocytes was assessed in the + hCSE group and normal medium (NM) groups（10% FBS）.
Results
The results of live/dead staining show a large number of live cells on the fresh lenticules. In the + CSE group, the CSCs had dendritic or stellate shapes under light microscopy; expressed keratocyte markers (ALDH3A1 and lumican), and were weakly stained with fibroblast and myofibroblast markers (fibronectin and α-SMA) via IF; showed that the keratocyte marker-related genes ALDH3A1, CD34, KERA and LUM were significantly greater and the fibrotic genes (FN1 and THBS1) were significantly lower than in the NM group via RT-qPCR analysis.
Conclusions
The CSCs were successfully isolated from SMILE-derived lenticules, and low-serum RIFA medium supplemented with hCSE can improve CSCs proliferation and antiapoptosis satisfactorily.