Characterization of kidney epithelial cells from the Florida manatee, Trichechus manatus latirostris

James M. Sweat, David D. Dunigan, Scott D. Wright

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The West-Indian manatee, Trichechus manatus latirostris, is a herbivorous marine mammal found in the coastal waters of Florida. Because of their endangered status, animal experimentation is not allowed. Therefore, a cell line was developed and characterized from tissue collected during necropsies of the manatees. A primary cell culture was established by isolating single cells from kidney tissue using both enzymatic and mechanical techniques. Primary manatee kidney (MK) cells were subcultured for characterization. These cells were morphologically similar to the cell lines of epithelial origin. An immunocytochemistry assay was used to localize the cytokeratin filaments common to cells of epithelial origin. At second passage, epithelial-like cells had an average population-doubling time of 48 h, had an optimum seeding density of 5 × 103 cells/cm2, and readily attached to plastic culture plates with a high level of seeding efficiency. Although the epithelial-like cells had a rapid growth rate during the first three passages, the cloning potential was low. These cells did not form colonies in agar medium, were serum dependent, had a limited life span of approximately nine passages, and possessed cell-contact inhibition. These data suggest that the cells were finite (noncontinuous growth), did not possess transformed properties, and were of epithelial origin. These cells are now referred to as MK epithelial cells.

Original languageEnglish (US)
Pages (from-to)386-394
Number of pages9
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume37
Issue number6
DOIs
StatePublished - Aug 29 2001

Fingerprint

Trichechus manatus
Epithelial Cells
Cells
Tissue
Kidney
Trichechus
Mammals
Cloning
Keratins
Cell culture
Agar
Assays
Animals
Plastics
Water
Contact Inhibition
Cell Line
Primary Cell Culture
Growth
Organism Cloning

Keywords

  • Cell culture
  • Cytokeratins
  • Endangered species
  • Florida manatee

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Characterization of kidney epithelial cells from the Florida manatee, Trichechus manatus latirostris. / Sweat, James M.; Dunigan, David D.; Wright, Scott D.

In: In Vitro Cellular and Developmental Biology - Animal, Vol. 37, No. 6, 29.08.2001, p. 386-394.

Research output: Contribution to journalArticle

@article{962c7459681e4d459f55a59830dd4957,
title = "Characterization of kidney epithelial cells from the Florida manatee, Trichechus manatus latirostris",
abstract = "The West-Indian manatee, Trichechus manatus latirostris, is a herbivorous marine mammal found in the coastal waters of Florida. Because of their endangered status, animal experimentation is not allowed. Therefore, a cell line was developed and characterized from tissue collected during necropsies of the manatees. A primary cell culture was established by isolating single cells from kidney tissue using both enzymatic and mechanical techniques. Primary manatee kidney (MK) cells were subcultured for characterization. These cells were morphologically similar to the cell lines of epithelial origin. An immunocytochemistry assay was used to localize the cytokeratin filaments common to cells of epithelial origin. At second passage, epithelial-like cells had an average population-doubling time of 48 h, had an optimum seeding density of 5 × 103 cells/cm2, and readily attached to plastic culture plates with a high level of seeding efficiency. Although the epithelial-like cells had a rapid growth rate during the first three passages, the cloning potential was low. These cells did not form colonies in agar medium, were serum dependent, had a limited life span of approximately nine passages, and possessed cell-contact inhibition. These data suggest that the cells were finite (noncontinuous growth), did not possess transformed properties, and were of epithelial origin. These cells are now referred to as MK epithelial cells.",
keywords = "Cell culture, Cytokeratins, Endangered species, Florida manatee",
author = "Sweat, {James M.} and Dunigan, {David D.} and Wright, {Scott D.}",
year = "2001",
month = "8",
day = "29",
doi = "10.1290/1071-2690(2001)037<0386:COKECF>2.0.CO;2",
language = "English (US)",
volume = "37",
pages = "386--394",
journal = "In Vitro Cellular and Developmental Biology - Plant",
issn = "1054-5476",
publisher = "Springer New York",
number = "6",

}

TY - JOUR

T1 - Characterization of kidney epithelial cells from the Florida manatee, Trichechus manatus latirostris

AU - Sweat, James M.

AU - Dunigan, David D.

AU - Wright, Scott D.

PY - 2001/8/29

Y1 - 2001/8/29

N2 - The West-Indian manatee, Trichechus manatus latirostris, is a herbivorous marine mammal found in the coastal waters of Florida. Because of their endangered status, animal experimentation is not allowed. Therefore, a cell line was developed and characterized from tissue collected during necropsies of the manatees. A primary cell culture was established by isolating single cells from kidney tissue using both enzymatic and mechanical techniques. Primary manatee kidney (MK) cells were subcultured for characterization. These cells were morphologically similar to the cell lines of epithelial origin. An immunocytochemistry assay was used to localize the cytokeratin filaments common to cells of epithelial origin. At second passage, epithelial-like cells had an average population-doubling time of 48 h, had an optimum seeding density of 5 × 103 cells/cm2, and readily attached to plastic culture plates with a high level of seeding efficiency. Although the epithelial-like cells had a rapid growth rate during the first three passages, the cloning potential was low. These cells did not form colonies in agar medium, were serum dependent, had a limited life span of approximately nine passages, and possessed cell-contact inhibition. These data suggest that the cells were finite (noncontinuous growth), did not possess transformed properties, and were of epithelial origin. These cells are now referred to as MK epithelial cells.

AB - The West-Indian manatee, Trichechus manatus latirostris, is a herbivorous marine mammal found in the coastal waters of Florida. Because of their endangered status, animal experimentation is not allowed. Therefore, a cell line was developed and characterized from tissue collected during necropsies of the manatees. A primary cell culture was established by isolating single cells from kidney tissue using both enzymatic and mechanical techniques. Primary manatee kidney (MK) cells were subcultured for characterization. These cells were morphologically similar to the cell lines of epithelial origin. An immunocytochemistry assay was used to localize the cytokeratin filaments common to cells of epithelial origin. At second passage, epithelial-like cells had an average population-doubling time of 48 h, had an optimum seeding density of 5 × 103 cells/cm2, and readily attached to plastic culture plates with a high level of seeding efficiency. Although the epithelial-like cells had a rapid growth rate during the first three passages, the cloning potential was low. These cells did not form colonies in agar medium, were serum dependent, had a limited life span of approximately nine passages, and possessed cell-contact inhibition. These data suggest that the cells were finite (noncontinuous growth), did not possess transformed properties, and were of epithelial origin. These cells are now referred to as MK epithelial cells.

KW - Cell culture

KW - Cytokeratins

KW - Endangered species

KW - Florida manatee

UR - http://www.scopus.com/inward/record.url?scp=0034878318&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034878318&partnerID=8YFLogxK

U2 - 10.1290/1071-2690(2001)037<0386:COKECF>2.0.CO;2

DO - 10.1290/1071-2690(2001)037<0386:COKECF>2.0.CO;2

M3 - Article

C2 - 11515973

AN - SCOPUS:0034878318

VL - 37

SP - 386

EP - 394

JO - In Vitro Cellular and Developmental Biology - Plant

JF - In Vitro Cellular and Developmental Biology - Plant

SN - 1054-5476

IS - 6

ER -