Mouse retina explants after long-term culture in serum free medium.
The neonatal mouse retina remains viable as an explant in serum-supplemented growth media for more than 4 weeks. Interpretation of drug effects on this tissue is compromised by the enigmatic composition of the serum. We sought to remove this ambiguity by culturing neonatal as well as late postnatal mouse retina in serum-free nutrient medium. In this study three important observations were made, (1) there is histotypic development of neonatal as well as preservation of late postnatal mouse retinal structure during long-term culture in serum-free medium, although the late postnatal tissue tends to show some loss of cells in the outer nuclear layer. (2) Protein expression in explant photoreceptor cells was similar to that in the litter-matched ones, except for green cone opsin and interphotoreceptor retinoid-binding protein, although mRNA of the latter is present at similar amounts as in age-matched in vivo controls. (3) Cells of the inner retina stained by antibodies to calcium-binding proteins display some novel sprouting of processes. The results show that the mouse retina can be cultured as an explant for more than 4 weeks in a serum-free medium. This represents an important step forward because, (1) the possibility of interference of drug effects by unknown serum factors has been eliminated; and (2) the spent culture medium can be analyzed to investigate biomolecules released by the retina in vitro.
Caffé
A R
AR
Department of Ophthalmology, Biomedical Center-Wallenberg Retina Center, University of Lund, Klinikgatan 26, S-221 84 Lund, Sweden.
Ahuja
P
P
Holmqvist
B
B
Azadi
S
S
Forsell
J
J
Holmqvist
I
I
Söderpalm
A K
AK
van Veen
T
T
D000818:Animals; D000906:Antibodies; D064032:Calbindin 2; D064026:Calbindins; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D016895:Culture Media, Serum-Free / Q000494:pharmacology*; D004801:Eosine Yellowish-(YS); D005136:Eye Proteins; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D006416:Hematoxylin; D051379:Mice; D010320:Parvalbumins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D017949:Retinal Cone Photoreceptor Cells / Q000737:chemistry / Q000166:cytology* / Q000276:immunology; D012177:Retinol-Binding Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D012243:Rhodopsin / Q000032:analysis / Q000276:immunology; D064030:S100 Calcium Binding Protein G / Q000032:analysis / Q000235:genetics / Q000276:immunology; D000099214:Retinol-Binding Proteins, Interstitial
D000906:Antibodies; D064032:Calbindin 2; D064026:Calbindins; D004396:Coloring Agents; D016895:Culture Media, Serum-Free; D004801:Eosine Yellowish-(YS); D005136:Eye Proteins; D006416:Hematoxylin; D010320:Parvalbumins; D012333:RNA, Messenger; D012177:Retinol-Binding Proteins; D012243:Rhodopsin; D064030:S100 Calcium Binding Protein G; D000099214:Retinol-Binding Proteins, Interstitial