Differentiation potential of human adipose tissue derived stem cells into photoreceptors through explants culture and enzyme methods.
OBJECTIVE
To investigate the retinal photoreceptor differentiation potential of human orbital adipose tissue-derived stem cells (ADSCs) generated by enzyme (EN) and explant (EX) culture methods.


METHODS
We investigated potentials of human orbital ADSCs to differentiate into photoreceptors through EN and EX culture methods. EN and EX orbital ADSCs were obtained from the same donor during rehabilitative orbital decompression, and then were subject to a 3-step induction using Noggin, DKK-1, IGF-1 and b-FGF at different time points for 38d. Stem cell, eye-field and photoreceptor-related gene and protein markers were measured by reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescent (IMF) staining.


RESULTS
Both EX and EN orbital ADSCs expressed CD133, a marker of cell differentiation. Moreover, PAX6 and rhodopsin, markers of the retinal progenitor cells, were detected from EX and EN orbital ADSCs. In EX orbital ADSCs, PAX6 mRNA was detected on the 17thday and then the rhodopsin mRNA was detected on the 24thday. In contrast, the EN orbital ADSCs expressed PAX6 and rhodopsin mRNA on the 31stday. EX orbital ADSCs expressed rhodopsin protein on the 24thday, while EN orbital ADSCs expressed rhodopsin protein on the 31stday.


CONCLUSIONS
Orbital ADSCs isolated by direct explants culture show earlier and stronger expressions of markers towards eye field and retinal photoreceptor differentiation than those generated by conventional EN method.
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong 999077, China; Department of Ophthalmology, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
Wei-Wei
WW
Xu
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong 999077, China.
Li
L
Huang
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong 999077, China.
Kelvin K L
KK
Chong
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong 999077, China.
Doreen S Y
DS
Leung
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong 999077, China.
Benjamin F L
BF
Li
Department of Ophthalmology, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
Zheng-Qin
ZQ
Yin
Department of Ophthalmology, Chinese People's Liberation Army General Hospital, Beijing 100853, China.
Yi-Fei
YF
Huang
Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong, Hong Kong 999077, China.
Chi Pui
CP
Pang
adult stem cells; cell differentiation; enzymatic digestion; explants culture; photoreceptor cells; tissue engineering