Characterization of rat colonic epithelial cell populations

Rabi Patnaik, Amitabha Mazumder, Belur Bhagavan, P. P. Nair

Research output: Contribution to journalArticle

Abstract

Colonic epithelial cells from male Sprague-Dawley rats were isolated by incubating everted colon with hyaluronidase suspended in Puck's saline F with an average cell yield of 120 × 106. These cells were fractionated by discontinuous Ficoll gradient and by short-term cell culture techniques. Centrifugation of isolated cells on discontinuous Ficoll gradient (15-35%) yielded populations differing in their proliferative activity. Additionally, a short-term cell culture technique was standardized to fractionate these cells according to their proliferating activity as judged by their DNA synthesis and thymidine kinase activity. Viability of these cells were judged by trypan blue exclusion, capacity to oxidize glucose and incorporation of precursors into protein DNA, RNA and glycoproteins. These fractionated cells were examined and identified by cytological studies. Cells showing proliferative activity sedimented at heavier regions of the Ficoll gradient, and the majority of these cells attached to the surface under conditions of short-term culture. Columnar mature absorptive cells and mucus-secreting goblet cells that showed very little proliferative activity sedimented at lighter zones of the Ficoll gradient and a major portion of these cells failed to attach by the cell culture method.

Original languageEnglish (US)
Pages (from-to)147-156
Number of pages10
JournalCell Differentiation
Volume10
Issue number3
DOIs
StatePublished - 1981

Fingerprint

Epithelial Cells
Ficoll
Population
Cell Culture Techniques
Polynucleotide 5'-Hydroxyl-Kinase
Hyaluronoglucosaminidase
Goblet Cells
Protein Precursors
Trypan Blue
Thymidine Kinase
Mucus
Centrifugation
Sprague Dawley Rats
Cell Survival
Glycoproteins
Colon
RNA
Glucose
DNA

Keywords

  • cell separation
  • colon epithelial cells
  • DNA synthesis

ASJC Scopus subject areas

  • Developmental Biology

Cite this

Characterization of rat colonic epithelial cell populations. / Patnaik, Rabi; Mazumder, Amitabha; Bhagavan, Belur; Nair, P. P.

In: Cell Differentiation, Vol. 10, No. 3, 1981, p. 147-156.

Research output: Contribution to journalArticle

Patnaik, Rabi ; Mazumder, Amitabha ; Bhagavan, Belur ; Nair, P. P. / Characterization of rat colonic epithelial cell populations. In: Cell Differentiation. 1981 ; Vol. 10, No. 3. pp. 147-156.
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