[Long-term expansion of retinal progenitor cells in vitro by culturing intact neurospheres in monolayer].
OBJECTIVE
To develop an efficient culture system for expansion of retinal progenitor cells for long-term in vitro.


METHODS
Retinal cells from embryonic mice were cultured as neurospheres-adhesive or as simple monolayer culture. For neurospheres-adhesive culture, spheres collected by low centrifugation from floating cultures were re-seeded onto Poly-D-Lysine coated flasks containing retinal progenitor cells(RPC) medium plus 10% FBS for the first 24 hours, then cultured in RPC medium (including FGF, EGF and LIF). For simple monolayer culture, on the other hand, cell suspension were directly plated on culture flasks pre-coated with Poly-D-Lysine. Proliferation and differentiation capability of the RPC from these two culture systems were analyzed by immunocytochemistry and FACS analysis.


RESULTS
Intact neurospheres-adhesive culture system was successful in expansion of RPC which maintain in uncommitted state. Immunochemistry staining and FACS analyzing demonstrated that the rate of nestin positive cells in uncommitted state (92%) and mature retinal neuron upon differential (53.7%) were higher than their counterparts in simple-adhesive cultures.


CONCLUSIONS
In this study we acquired a stable expansion of RPC which preferentially differentiate into mature retinal neurons.
Lai
Pinghong
P
Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China.
Tang
Shibo
S
Lin
Shaofen
S
Zhu
Xiaobo
X
Gao
Yi
Y
Meng
Jing
J
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D049109:Cell Proliferation; D051379:Mice; D012160:Retina / Q000166:cytology* / Q000196:embryology; D013234:Stem Cells / Q000166:cytology*