The effects of human serum to the morphology, proliferation and gene expression level of the respiratory epithelium in vitro

Mohd Heikal Mohd Yunus, Kan Chan Siang, Nurul Izzati Hashim, Ng Pei Zhi, Nur Fathurah Zamani, Primuharsa Putra Sabri, Mohd Fauzi Busra, Shiplu Roy Chowdhury, Ruszymah Binti Haji Idrus

Research output: Contribution to journalArticle

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Abstract

The culture of human airway epithelial cells has played an important role in advancing our understanding of the metabolic and molecular mechanisms underlying normal function and disease pathology of airway epithelial cells. The present study focused on investigating the effects of human serum (HS) on the qualitative and quantitative properties of the human respiratory epithelium compared to the fetal bovine serum (FBS), as a supplement in culture. Respiratory epithelial (RE) cells derived from human nasal turbinate were co-cultured with fibroblasts, subsequently separated at 80-90% confluency by differential trypsinization. RE cells were then sub-cultured into 2 different plates containing 5% allogenic HS and FBS supplemented media respectively up to passage 1 (P1). Cell morphology, growth rate, cell viability and population doubling time were assessed under light microscope, and levels of gene expression were measured via real time reverse transcriptase-polymerase chain reaction (qRT-PCR). RE cells appeared as polygonal shape and expanded when cultured in HS whereas RE cells in FBS were observed to be easily matured thus limit the RE cells expansion. Proliferation rate of RE cells in HS supplemented media (7673.18±1207.15) was 3 times higher compared to RE in FBS supplemented media (2357.68±186.85). Furthermore, RE cells cultured in HS-supplemented media required fewer days (9.15±1.10) to double in numbers compared to cells cultured in FBS-supplemented media (13.66±0.81). Both the differences were significant (p<0.05). However, there were no significant differences in the viability of RE cells in both groups (p=0.105). qRT-PCR showed comparable expressions of gene Cytokeratin-14 (CK-14), Cytokeratin-18 (CK-18) and Mucin-5 subtype B (MUC5B) in RE cells cultured in both groups (p>0.05). In conclusion, HS is a comparatively better choice of media supplement in accelerating growth kinetics of RE cells in vitro thus producing a better quality of respiratory epithelium for future tracheal reconstruction.

Original languageEnglish
Pages (from-to)233-240
Number of pages8
JournalTissue and Cell
Volume46
Issue number4
DOIs
Publication statusPublished - 2014

Fingerprint

Respiratory Mucosa
Epithelial Cells
Gene Expression
Serum
In Vitro Techniques
Turbinates
Respiratory Rate
Growth
Reverse Transcriptase Polymerase Chain Reaction
Nose
Real-Time Polymerase Chain Reaction
Cultured Cells
Cell Survival
Fibroblasts

Keywords

  • Fetal bovine serum
  • Gene expression
  • Growth kinetics
  • Human serum
  • Respiratory epithelium

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology

Cite this

The effects of human serum to the morphology, proliferation and gene expression level of the respiratory epithelium in vitro. / Mohd Yunus, Mohd Heikal; Chan Siang, Kan; Hashim, Nurul Izzati; Pei Zhi, Ng; Zamani, Nur Fathurah; Putra Sabri, Primuharsa; Busra, Mohd Fauzi; Roy Chowdhury, Shiplu; Binti Haji Idrus, Ruszymah.

In: Tissue and Cell, Vol. 46, No. 4, 2014, p. 233-240.

Research output: Contribution to journalArticle

Mohd Yunus, Mohd Heikal ; Chan Siang, Kan ; Hashim, Nurul Izzati ; Pei Zhi, Ng ; Zamani, Nur Fathurah ; Putra Sabri, Primuharsa ; Busra, Mohd Fauzi ; Roy Chowdhury, Shiplu ; Binti Haji Idrus, Ruszymah. / The effects of human serum to the morphology, proliferation and gene expression level of the respiratory epithelium in vitro. In: Tissue and Cell. 2014 ; Vol. 46, No. 4. pp. 233-240.
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AU - Pei Zhi, Ng

AU - Zamani, Nur Fathurah

AU - Putra Sabri, Primuharsa

AU - Busra, Mohd Fauzi

AU - Roy Chowdhury, Shiplu

AU - Binti Haji Idrus, Ruszymah

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