Use of human serum for human corneal endothelial cell culture

Abstract

HS-SM was similar to FBS-SM for HCEC culture when assessed by cell morphology, proliferation and protein/gene expression.

Transcriptomic Analysis of Cultured Corneal Endothelial Cells as a Validation for Their Use in Cell Replacement Therapy

“…Identifying the growth conditions that support the growth of cells with a transcriptome profile similar to evHCEnC will provide researchers with a more accurate model of in vivo HCEnC and a potential source of endothelial cells for management of endothelial dysfunction. Optimization of HCEnC culturing techniques should take into consideration several anatomic and physiologic features of in vivo HCEnC (60), such as: 1) adherence to a complex milieu of extracellular matrix proteins (13,18,22,38,40,51,57); 2) contact with physiological proteins and other factors present in aqueous humor (25,33,36,42,53); 3) exposure to appropriate biomechanical forces (35,40,57); and 4) maintenance of a confluent semipermeable layer of cells with strong apicobasal polarity. Replication of each of these features in a single culturing method would be challenging, but would represent a significant advance in the development of cultured HCEnC that closely resemble in vivo HCEnC.…”

Characterization of cryopreserved primary human corneal endothelial cells cultured in human serum-supplemented media

“…Unpublished observations during our previous study (9) have demonstrated that cells grown in HS-SM detach faster than those grown in FBS-SM. Similarly, in this study, we observed a tendency of longer detachment time with FBS-SM and lower passages.…”

“…The development in surgical treatment of corneal endothelial diseases, with progressive thinning of grafts and its related advantages (13,14) , leads us to conclude that transplantation with cultured en- dothelial cells may represent the next advance in this field (2)(3)(4)(5)(6)(7)(8)(9) . Animal studies have demonstrated promising results (15,16) .…”

“…culture media FBS-SM was previously described in a study (12) and was used in our laboratory (11) and comprised serum-free medium (OptiMEM-I), 8% FBS, 5 ng/mL hEGF, 20 ng/mL NGF, 100 mcg/mL BPE, 20 mcg/mL ascorbic acid, 200 mg/L calcium chloride, 0.08% chondroitin sulfate, 50 mcg/mL gentamicin, and antibiotic/antimycotic solution (diluted 1/100). HS-SM was described in our previous study (9) and comprised the same components as FBS-SM, with the exception that 8% FBS was replaced with 8% HS. The concentration of serum (both in FBS and HS) that was used to culture cells in glass chamber slides was doubled to 16%.…”

“…Regarding the first area, we recently published an article demonstrating the applicability of human serum-supplemented medium (HS-SM) as an alternative to fetal bovine serum-supplemented media (FBS-SM), suggesting potential advantages of a medium with fewer animal-derived components (9) . Regarding the second area, the cryo preservation of both corneas for transplantation (10) and cultu red HCEC using a standard medium (FBS-SM) have been already demons trated (11) and could potentially improve the logistics related to the distribution of cultured HCECs for clinical use.…”