Stem cell genes are poorly expressed in chondrocytes from microtic cartilage

M. F. Ishak, Chua Kien Hui, Asma Abdullah, L. Saim, B. S. Aminuddin, Ruszymah Idrus, Bee See Goh

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Objectives: This study was aimed to see the difference between chondrocytes from normal cartilage compared to chondrocytes from microtic cartilage. Specific attentions were to characterize the growth of chondrocytes in terms of cell morphology, growth profile and RT-PCR analysis. Study design: Laboratory experiment using auricular chondrocytes. Methods: Chondrocytes were isolated from normal and microtic human auricular cartilage after ear reconstructive surgeries carried out at the Universiti Kebangsaan Malaysia Medical Centre. Chondrocytes were cultured in vitro and subcultured until passage 4. Upon confluency, cultured chondrocytes at each passage (P1, P2, P3 and P4) were harvested and subjected to growth profile and gene expression analyses. Comparison was made between the microtic and normal chondrocytes. Results: For growth profile analysis cell viability did not show significant differences between both samples. There are no significance differences between both samples in terms of its growth rate, except in passage 1 where microtic chondrocytes were significant lower in their growth rate. Population doubling time and total number of cell doubling of all samples also did not show any significant differences. Gene expression is measured using Real Time-Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). There is no significant differences in the expression of collagen type I, collagen type II, collagen type X, aggrecan core protein, elastin and sox9 genes in both samples. There are significant lower in the expression of sox2, nestin, BST-1 and OCT-4 gene in microtic chondrocytes compared to the normal chondrocytes. Stem cells markers are included in this study as stemness in cells may imply a greater proliferative potential and plasticity in vitro. Conclusion: Chondrocytes from microtic samples have the same properties as chondrocytes from normal samples and hold promises to be used as a starting material in the reconstruction of the external ear in future clinical application. The reduction in sox2, nestin, BST-1 and OCT-4 gene expression in microtic samples could be the possible cause of the arrested development of the external ear.

Original languageEnglish
Pages (from-to)835-840
Number of pages6
JournalInternational Journal of Pediatric Otorhinolaryngology
Volume75
Issue number6
DOIs
Publication statusPublished - Jun 2011

Fingerprint

Chondrocytes
Cartilage
Stem Cells
Genes
Growth
External Ear
Nestin
Reverse Transcriptase Polymerase Chain Reaction
Reconstructive Surgical Procedures
Collagen Type X
Ear Cartilage
Gene Expression
Aggrecans
Collagen Type II
Elastin
Malaysia
Collagen Type I
Transcriptome
Ear
Cell Survival

Keywords

  • Auricular chondrocytes
  • Microtia
  • Stem cell
  • Tissue engineering

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Pediatrics, Perinatology, and Child Health

Cite this

Stem cell genes are poorly expressed in chondrocytes from microtic cartilage. / Ishak, M. F.; Kien Hui, Chua; Abdullah, Asma; Saim, L.; Aminuddin, B. S.; Idrus, Ruszymah; Goh, Bee See.

In: International Journal of Pediatric Otorhinolaryngology, Vol. 75, No. 6, 06.2011, p. 835-840.

Research output: Contribution to journalArticle

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AU - Abdullah, Asma

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AU - Aminuddin, B. S.

AU - Idrus, Ruszymah

AU - Goh, Bee See

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AB - Objectives: This study was aimed to see the difference between chondrocytes from normal cartilage compared to chondrocytes from microtic cartilage. Specific attentions were to characterize the growth of chondrocytes in terms of cell morphology, growth profile and RT-PCR analysis. Study design: Laboratory experiment using auricular chondrocytes. Methods: Chondrocytes were isolated from normal and microtic human auricular cartilage after ear reconstructive surgeries carried out at the Universiti Kebangsaan Malaysia Medical Centre. Chondrocytes were cultured in vitro and subcultured until passage 4. Upon confluency, cultured chondrocytes at each passage (P1, P2, P3 and P4) were harvested and subjected to growth profile and gene expression analyses. Comparison was made between the microtic and normal chondrocytes. Results: For growth profile analysis cell viability did not show significant differences between both samples. There are no significance differences between both samples in terms of its growth rate, except in passage 1 where microtic chondrocytes were significant lower in their growth rate. Population doubling time and total number of cell doubling of all samples also did not show any significant differences. Gene expression is measured using Real Time-Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). There is no significant differences in the expression of collagen type I, collagen type II, collagen type X, aggrecan core protein, elastin and sox9 genes in both samples. There are significant lower in the expression of sox2, nestin, BST-1 and OCT-4 gene in microtic chondrocytes compared to the normal chondrocytes. Stem cells markers are included in this study as stemness in cells may imply a greater proliferative potential and plasticity in vitro. Conclusion: Chondrocytes from microtic samples have the same properties as chondrocytes from normal samples and hold promises to be used as a starting material in the reconstruction of the external ear in future clinical application. The reduction in sox2, nestin, BST-1 and OCT-4 gene expression in microtic samples could be the possible cause of the arrested development of the external ear.

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KW - Microtia

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