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Tytuł pozycji:

Development-related mitochondrial properties of retinal pigment epithelium cells derived from hEROs

Tytuł:
Development-related mitochondrial properties of retinal pigment epithelium cells derived from hEROs
Autorzy:
Hao-Jue Xu
Qi-You Li
Ting Zou
Zheng-Qin Yin
Temat:
mitochondria
retinal organoids
retinal pigment epithelium cells
human embryonic stem cells
retinal degenerative diseases
Ophthalmology
RE1-994
Źródło:
International Journal of Ophthalmology, Vol 14, Iss 8, Pp 1138-1150 (2021)
Wydawca:
Press of International Journal of Ophthalmology (IJO PRESS), 2021.
Rok publikacji:
2021
Kolekcja:
LCC:Ophthalmology
Typ dokumentu:
article
Opis pliku:
electronic resource
Język:
English
ISSN:
2222-3959
2227-4898
Relacje:
http://ies.ijo.cn/en_publish/2021/8/20210802.pdf; https://doaj.org/toc/2222-3959; https://doaj.org/toc/2227-4898
DOI:
10.18240/ijo.2021.08.02
Dostęp URL:
https://doaj.org/article/2162b567c2db4665a40c4ffe92f78ffd  Link otwiera się w nowym oknie
Numer akcesji:
edsdoj.2162b567c2db4665a40c4ffe92f78ffd
Czasopismo naukowe
AIM: To explore the temporal mitochondrial characteristics of retinal pigment epithelium (RPE) cells obtained from human embryonic stem cells (hESC)-derived retinal organoids (hEROs-RPE), to verify the optimal period for using hEROs-RPE as donor cells from the aspect of mitochondria and to optimize RPE cell-based therapeutic strategies for age-related macular degeneration (AMD). METHODS: RPE cells were obtained from hEROs and from spontaneous differentiation (SD-RPE). The mitochondrial characteristics were analyzed every 20d from day 60 to 160. Mitochondrial quantity was measured by MitoTracker Green staining. Transmission electron microscopy (TEM) was adopted to assess the morphological features of the mitochondria, including their distribution, length, and cristae. Mitochondrial membrane potentials (MMPs) were determined by JC-1 staining and evaluated by flow cytometry, reactive oxygen species (ROS) levels were evaluated by flow cytometry, and adenosine triphosphate (ATP) levels were measured by a luminometer. Differences between two groups were analyzed by the independent-samples t-test, and comparisons among multiple groups were made using one-way ANOVA or Kruskal-Wallis H test when equal variance was not assumed. RESULTS: hEROs-RPE and SD-RPE cells from day 60 to 160 were successfully differentiated from hESCs and expressed RPE markers (Pax6, MITF, Bestrophin-1, RPE65, Cralbp). RPE features, including a cobblestone-like morphology with tight junctions (ZO-1), pigments and microvilli, were also observed in both hEROs-RPE and SD-RPE cells. The mitochondrial quantities of hEROs-RPE and SD-RPE cells both peaked at day 80. However, the cristae of hEROs-RPE mitochondria were less mature and abundant than those of SD-RPE mitochondria at day 80, with hEROs-RPE mitochondria becoming mature at day 100. Both hEROs-RPE and SD-RPE cells showed low ROS levels from day 100 to 140 and maintained a normal MMP during this period. However, hEROs-RPE mitochondria maintained a longer time to produce high levels of ATP (from day 120 to 140) than SD-RPE cells (only day 120). CONCLUSION: hEROs-RPE mitochondria develop more slowly and maintain a longer time to supply high-level energy than SD-RPE mitochondria. From the mitochondrial perspective, hEROs-RPE cells from day 100 to 140 are an optimal cell source for treating AMD.

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