title: The effects of human serum to the morphology, proliferation and gene expression level of the respiratory epithelium in vitro.
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.
authors: Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia. Electronic address: mohdheikalyunus@yahoo.com.
authors: Mohd Heikal Mohd
authors: MH
authors: Yunus
authors: Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Kan Chan
authors: KC
authors: Siang
authors: Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Nurul Izzati
authors: NI
authors: Hashim
authors: Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Ng Pei
authors: NP
authors: Zhi
authors: Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Nur Fathurah
authors: NF
authors: Zamani
authors: KPJ Seremban Specialist Hospital, Negeri Sembilan, Malaysia.
authors: Primuharsa Putra
authors: PP
authors: Sabri
authors: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Mohd Fauzi
authors: MF
authors: Busra
authors: Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Shiplu Roy
authors: SR
authors: Chowdhury
authors: Department of Physiology, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia; Tissue Engineering Centre, Universiti Kebangsaan Malaysia Medical Centre, Kuala Lumpur, Malaysia.
authors: Ruszymah Binti Haji
authors: RB
authors: Idrus
mesh_terms: D000818:Animals; D002417:Cattle; D049109:Cell Proliferation; D018920:Coculture Techniques; D005786:Gene Expression Regulation; D006801:Humans; D066298:In Vitro Techniques; D053547:Keratin-14 / Q000096:biosynthesis; D053538:Keratin-18 / Q000096:biosynthesis; D055257:Mucin-5B / Q000096:biosynthesis; D061251:Primary Cell Culture; D020545:Respiratory Mucosa / Q000166:cytology*; D044967:Serum / Q000737:chemistry / Q000378:metabolism*
chemical_list: D053547:Keratin-14; D053538:Keratin-18; D055257:Mucin-5B
keywords: Fetal bovine serum; Gene expression; Growth kinetics; Human serum; Respiratory epithelium