Characterization of cultivated murine lacrimal gland epithelial cells.
OBJECTIVE
To date, mouse lacrimal gland epithelial cells have been cultured successfully but only in cases involving newborn mouse lacrimal glands. In this work, we attempted to cultivate and characterize adult mouse lacrimal gland epithelial cells.


METHODS
Lacrimal glands were removed from newborn mice (C57B/6) and isolated lacrimal gland epithelial cells were seeded onto tissue culture treated or low adherent culture dishes in Cnt-07 culture medium with or without cholera toxin. Cultivated cells were characterized by immunostaining with pan-cytokeratin, α-smooth muscle actin, and lactoferrin antibodies. Lacrimal gland cells from 7-week-old green fluorescent protein (GFP) and non-GFP (C57B/6) mice were mixed and seeded onto uncoated dishes to assess sphere-forming efficiency. Cells were also seeded onto 3T3 cell feeder layers to assess colony forming efficiency.


RESULTS
Lacrimal gland epithelial cells were selectively cultured with cholera toxin, and cell type was verified by pan-cytokeratin and α-smooth muscle actin immunostaining. Sphere formation from single cells of adult mice was observed using specific medium and low adherent culture dishes. These cells could also undergo colony formation on 3T3 feeder cells.


CONCLUSIONS
Adult mouse lacrimal gland epithelial cells were successfully cultivated in cholera toxin-containing medium, and were observed to form spheres from single cells.
Department of Ophthalmology, Keio University School of Medicine, Tokyo, Japan.
Shinya
S
Kobayashi
Tetsuya
T
Kawakita
Motoko
M
Kawashima
Naoko
N
Okada
Kenji
K
Mishima
Ichiro
I
Saito
Masataka
M
Ito
Shigeto
S
Shimmura
Kazuo
K
Tsubota
D000199:Actins / Q000096:biosynthesis; D000818:Animals; D000831:Animals, Newborn; D041702:BALB 3T3 Cells; D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D002772:Cholera Toxin / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D061252:Feeder Cells / Q000166:cytology; D007633:Keratins / Q000096:biosynthesis; D007765:Lacrimal Apparatus / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007781:Lactoferrin / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL
D000199:Actins; C544013:alpha-smooth muscle actin, mouse; D007633:Keratins; D002772:Cholera Toxin; D007781:Lactoferrin