Growth and differentiation of the cultured secretory cells of the cow oviduct on reconstituted basement membrane

Abstract

Isolated bovine oviduct epithelial cells were cultured on plastic precoated with matrigel. The epithelial cells seeded on 10 mg/ml matrigel often organized themselves into hollow tubes or spheres with microvilli directed towards the lumen. This is the first report of describing the spontaneous tube formation of oviduct epithelial cells in vitro. The epithelial cells growing on this substratum became fully differentiated with the formation of junctional complexes and the production of secretory vesicles which migrated to apical plasmalemma. Epithelial cells seeded on 2 mg/ml matrigel (dry film) formed a subconfluent monolayer in 15-20 days after seeding. The histoarchitecture of the secretory cells growing on the matrigel dry film closely resembled that of intact epithelial cells. Occasional ciliated cells containing large numbers of mitochondria were observed in cell colonies growing on 2 mg/ml matrigel but they possessed very few intact cilia. The monolayer linearly incorporated 35S-methionine into proteins up to 8 hr in presence of estradiol or progesterone. The fluorography of the newly synthesized proteins indicated that the cell extracts of estradiol-stimulated cells contained an additional protein of approximate molecular weight of 60 kd as compared to the extracts of cells incubated without steroids or incubated with progesterone.

Morphological and functional characterization of bovine oviductal epithelial cell monolayers cultured on polarizing membranes

“…Cells cultured on inserts formed a confluent monolayer in 2-3 days, in contrast to that reported when other growth substrates are used (15-20 days in matrigel, Joshi, 1991) and by using non-polarizing conditions in 4-well plates (this work, 5-7 days) and in different culture supports (6 days in flasks, multiwell plates, culture dishes and glass coverslips, Van Langendonckt et al, 1995). The morphology of cells cultured in 4-well plates is in accordance with that previously described by Van (Lodish et al, 1986 (Leese and Gray, 1985).…”

“…Limited efforts to improve in vitro embryo production have been focused on cell-polarizing in co-cultures, possibly because high levels of cell polarization were not required for the beneficial effects of coculture to occur (Ouhibi et al, 1990;McCaffrey et al, 1991). Polarized cultured BOEC on matrigel coating showed morphological and functional features resembling those of intact epithelial cells (Ouhibi et al, 1990;McCaffrey et al, 1991;Eyestone et al, 1991;Joshi, 1991) even with spontaneous tubeshaped organization (Joshi, 1991) and a yield of blastocysts, co-cultured on frozen/thawed cell-monolayers, exhibiting a number of cells higher than that from nonpolarized cultured BOEC (Da Silva, 1994). Spermatozoa were able to bind more efficiently to polarized BOEC than to the same non-polarized cells (Pollard et al, 1991;Ellington et al, 1993 (Eyestone et al, 1991;Mermillod et al, 1993) (experiment 1 Some other samples were utilized fresh for pH determination or frozen/thawed for lactate determination (experiment 2).…”

Induction of Thumbtack Sperm During Coculture with Oviduct Epithelial Cell Monolayers in a Marsupial, the Brushtail Possum (Trichosurus vulpecula)1

“…Matrigel was found to help maintain the differentiated state of the epithelial cells for 7-10 days in brushtail possum. This observation is consistent with previous studies, in which Matrigel improved polarization and differentiation in cultured mammary epithelial cells [26], human endothelial cells [27], and bovine and equine OEC [24,28].…”

“…Evidence for monolayer viability was vigorous ciliary activity. The purity of epithelial cells was assessed using indirect immunofluorescence analysis of epithelial cytokeratins [24].…”

Collagen Vitrigel: A Novel Scaffold that can Facilitate a Three-Dimensional Culture for Reconstructing Organoids

“…Culture systems to reorganize a threebiomedical research. Human umbilical vein endothelial dimensional multicellular mass are classified into the folcells (HUVECs) cultured on matrigel formed a network lowing five categories: 1) gel culture systems utilizing of capillary-like structures (19) and human microvascucollagen gel and matrigel scaffolds (1,3-5, 11,[13][14][15]18, lar endothelial cells (HMVECs) also formed a network 19,23,28,29,39); 2) spheroid culture systems utilizing of branching tubular capillary-like structures into an thermo-responsive polymer, agarose, and polyurethane overlaid collagen gel with embedded fibroblasts (39), foam scaffolds (7,9,17,21,24,31-34); 3) medium circudemonstrating the reconstruction of in vitro angiogenlation culture systems utilizing scaffolds such as hollow esis models. The hetero-spheroids composed of mesenfiber and cotton gauze (12,25,35); 4) tissue architecturechymal cells (e.g., human dermal fibroblasts) and epi- thelial cells such as rat primary hepatocytes (32), human glass-like material and succeeded in preparing a novel, stable state of collagen gel possessing valuable physical epidermal keratinocytes (34), and human carcinoma cell lines (17,24) were prepared by the following procedure.…”