Chapter 10 Improved methodology for cell culture of human and monkey retinal pigment epithelium

Longterm cultures of the aged human RPE do not maintain epithelial morphology and high transepithelial resistance

“…In agreement with our observations at long-term PC, McKay and Pfeffer described phenotypic heterogeneity patterns with aRPE cell morphologies ranging from polygonal to spindle-shaped [5,6]. Comparable studies using fetal human [2], chick [7], rat RPE [8], or the ARPE19 cell line [3] show the influence of culture conditions on long-term differentiation.…”

Longterm cultures of the aged human RPE do not maintain epithelial morphology and high transepithelial resistance

“…In agreement with our observations at long-term PC, McKay and Pfeffer described phenotypic heterogeneity patterns with aRPE cell morphologies ranging from polygonal to spindle-shaped [5,6]. Comparable studies using fetal human [2], chick [7], rat RPE [8], or the ARPE19 cell line [3] show the influence of culture conditions on long-term differentiation.…”

Novel Enzymatic Isolation of an Entire Viable Human Limbal Epithelial Sheet

“…Because it was necessary to incubate in dispase II for such a long period, it is important to keep it at a low temperature (4°C) to reduce metabolic activity and maintain the tissue in a medium with growth supplements to maintain the viability and in the presence of 100 mM sorbitol to prevent cell swelling by increasing the osmolarity. 27 By doing so, we confirmed that isolated limbal epithelial sheets indeed retained a high viability of 80.7%. Because human limbal rings used in this study were not fresh and were studied at variable times after death, stored in Chen medium, and transported to the laboratory, there may have been cell death before digestion.…”

Cell culture conditions affect RPE phagocytic function

“…In 1990, Pfeffer described that retinal extract (RE) promotes RPE cell differentiation in vitro [42]. Our data show that RE causes an increase of phagocytosis at a concentration of 1%, whereas higher concentrations reduced that response significantly below control.…”

Longterm cultures of the aged human RPE do not maintain epithelial morphology and high transepithelial resistance

“…In agreement with our observations at long-term PC, McKay and Pfeffer described phenotypic heterogeneity patterns with aRPE cell morphologies ranging from polygonal to spindle-shaped [5,6]. Comparable studies using fetal human [2], chick [7], rat RPE [8], or the ARPE19 cell line [3] show the influence of culture conditions on long-term differentiation.…”

Novel Enzymatic Isolation of an Entire Viable Human Limbal Epithelial Sheet

“…Because it was necessary to incubate in dispase II for such a long period, it is important to keep it at a low temperature (4°C) to reduce metabolic activity and maintain the tissue in a medium with growth supplements to maintain the viability and in the presence of 100 mM sorbitol to prevent cell swelling by increasing the osmolarity. 27 By doing so, we confirmed that isolated limbal epithelial sheets indeed retained a high viability of 80.7%. Because human limbal rings used in this study were not fresh and were studied at variable times after death, stored in Chen medium, and transported to the laboratory, there may have been cell death before digestion.…”

Cell culture conditions affect RPE phagocytic function

“…In 1990, Pfeffer described that retinal extract (RE) promotes RPE cell differentiation in vitro [42]. Our data show that RE causes an increase of phagocytosis at a concentration of 1%, whereas higher concentrations reduced that response significantly below control.…”

Longterm cultures of the aged human RPE do not maintain epithelial morphology and high transepithelial resistance

“…In agreement with our observations at long-term PC, McKay and Pfeffer described phenotypic heterogeneity patterns with aRPE cell morphologies ranging from polygonal to spindle-shaped [5,6]. Comparable studies using fetal human [2], chick [7], rat RPE [8], or the ARPE19 cell line [3] show the influence of culture conditions on long-term differentiation.…”

Novel Enzymatic Isolation of an Entire Viable Human Limbal Epithelial Sheet

“…Because it was necessary to incubate in dispase II for such a long period, it is important to keep it at a low temperature (4°C) to reduce metabolic activity and maintain the tissue in a medium with growth supplements to maintain the viability and in the presence of 100 mM sorbitol to prevent cell swelling by increasing the osmolarity. 27 By doing so, we confirmed that isolated limbal epithelial sheets indeed retained a high viability of 80.7%. Because human limbal rings used in this study were not fresh and were studied at variable times after death, stored in Chen medium, and transported to the laboratory, there may have been cell death before digestion.…”

Cell culture conditions affect RPE phagocytic function

“…In 1990, Pfeffer described that retinal extract (RE) promotes RPE cell differentiation in vitro [42]. Our data show that RE causes an increase of phagocytosis at a concentration of 1%, whereas higher concentrations reduced that response significantly below control.…”