Nylon-screen filtration devices for crude separation of dissociated cells for primary culture

McAteer, James A.; Cavanagh, Thomas J.

doi:10.1007/bf01834635

Cited by 3 publications

(2 citation statements)

References 3 publications

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“…Repeat this step. Note: At this point it may be useful, depending on the specific application for the cell preparation, to filter the cell suspension through size-bruiting screen such as Nitex cloth (11). Exercise caution, because too fine a mesh may retain a substantial amount of the cell yield, particularly if polymerized DNA from lysed cells has not been diluted out.…”

Section: Methodsmentioning

confidence: 98%

Culture of human renal cortex epithelial cells

McAteer

Kempson

Evan

1991

Journal of Tissue Culture Methods

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SUMMARY: A procedure is described for the establishment and propagation of epithelial cell rich cultures derived from normal human kidney cortex (NHK-C cells). Ceils are harvested from tissue fragments of donor human kidney by progressive enzymatic dissociation. NHK-C cultures are morphologically heterogeneous but exhibit, predominantly, the functional characteristics of cells of the kidney proximal tubule.

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Section: Methodsmentioning

confidence: 98%

Culture of human renal cortex epithelial cells

McAteer

Kempson

Evan

1991

Journal of Tissue Culture Methods

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“…The cells were trypsinized and then washed twice in medium containing 0.1% soybean trypsin inhibitor (Boehringer-Mannheim, Indianapolis, IN). Cells were resuspended in culture medium to give 2.5 x 105 cells/ml and then gently triturated through an 18-gauge cannula and filtered (McAteer and Cavanagh, 1983) through 20-pm Nitex screen (Tetko Inc., Elmsford, NY). The dispersed cell suspension (4.3 ml) was added to a chilled collagen-medium solution.…”

Section: Culture Of Mdck Cells Suspended Within Collagen Gelmentioning

confidence: 99%

Morphogenetic clonal growth of kidney epithelial cell line MDCK

1987

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MDCK (Madin-Darby canine kidney) cells were cultured either 1) dispersed within hydrated collagen gel (HCG) or 2) seeded atop a collagen substrate and then immediately overlaid with HCG. Individual cells exhibited clonal growth in three dimensions to form spherical cysts made up of a simple epithelium enclosing a fluid-filled lumen. The cells of MDCK cysts were polarized with the basolateral surface in contact with the collagen gel and the apical surface bordering the lumen. The ultrastructure of MDCK cysts showed similarities to distal nephron. The cells bore apical microvilli and solitary cilia and had occluding junctions and a simple basolateral surface. MDCK cysts increased in size (> 800 pm diameter) with continued culture. MDCK cysts grown between layers of HCG were stripped free of the overlying collagen to give direct access to basolateral surface membrane. Unlike monolayer culture, morphogenetic clonal growth of cell line MDCK produces a polarized cell population with a true lumenal and basolateral surface. Collagen-gelcultured MDCK cysts provide an easily manipulable in uitro cell system that may offer unique advantages for the study of renal cell structure and function.

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MDCK cysts: An in vitro model of epithelial cyst formation and growth

McAteer

Evan

Vance

et al. 1986

Journal of Tissue Culture Methods

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SUMMARY: MDCK cells dissociated from monolayer culture were either dispersed within medium-hydrated collagen gel or seeded atop a collagen substrate which was immediately overlaid with collagen gel. Individual cells exhibited clonal growth in three dimensions to form spherical cysts in which a simple epithelium surrounded a fluid-filled lumen. The ceils of MDCK cysts were polarized with apical surface bordering the lumen. MDCK cysts increased in diameter with continued culture. Maximum cyst size was dependent on seeding density and was influenced by medium composition. MDCK cysts could be isolated from the collagen substrate by digestion with collagenase. Also, collagen gel could be dissected from the cyst wall to give unrestricted access to regions of the basolateral cell surface. This novel method of renal cell culture provides a study system to model the influence of the extracellular matrix on kidney epithelial cell structure and function. It also offers an in vitro model of general application to the study of epithelial cyst formation and growth.

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Nylon-screen filtration devices for crude separation of dissociated cells for primary culture

Cited by 3 publications

References 3 publications

Culture of human renal cortex epithelial cells

Culture of human renal cortex epithelial cells

Morphogenetic clonal growth of kidney epithelial cell line MDCK

MDCK cysts: An in vitro model of epithelial cyst formation and growth

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