Determination of the differentiation capacities of murines' primary mononucleated cells and MC3T3-E1 cells

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Background: The main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus. The main purpose of this study was to determine the differentiation capacities of stem and progenitor cells. This study was performed using mononucleated cells originated from murine peripheral blood and MC3T3-E1 cells. Three approaches were used to determine their differentiation capacities: 1) Biochemical assays, 2) Gene expression analysis, and 3) Morphological observations.Results: We found that both cells were able to differentiate into mature osteoblasts, as assayed by ALP activity. RT-PCR analysis showed the activation of the Opn gene after osteoblast differentiation. Morphological observations of both cells revealed the formation of black or dark-brown nodules after von Kossa staining. Nevertheless, only mononucleated cells showed the significant increase in TRAP activity characteristic of mature osteoclasts. The osteoclast-specific CatK gene was only upregulated in mononucleated cells. Morphological observations indicated the existence of multinucleated osteoclasts. Sca-1 was activated only in undifferentiated mononucleated cells, indicating that the cells were hematopoietic stem cells. In both cell lines, the housekeeping Gapdh gene was activated before and after differentiation.Conclusion: The isolated mononucleated cells were able to differentiate into both osteoblasts and osteoclasts; indicating that they are stem cells. On the other hand, MC3T3-E1 cells can only differentiate into osteoblasts; a characteristic of progenitor cells.

Original languageEnglish
Article number42
JournalCancer Cell International
Volume10
DOIs
Publication statusPublished - 28 Oct 2010

Fingerprint

Stem Cells
Osteoclasts
Osteoblasts
Activation Analysis
Essential Genes
Hematopoietic Stem Cells
Transcriptional Activation
Staining and Labeling
Gene Expression
Cell Line
Polymerase Chain Reaction
Genes

ASJC Scopus subject areas

  • Cancer Research
  • Oncology
  • Genetics

Cite this

@article{10497ac0cb9244a4b2ab5de4c2c9ace5,
title = "Determination of the differentiation capacities of murines' primary mononucleated cells and MC3T3-E1 cells",
abstract = "Background: The main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus. The main purpose of this study was to determine the differentiation capacities of stem and progenitor cells. This study was performed using mononucleated cells originated from murine peripheral blood and MC3T3-E1 cells. Three approaches were used to determine their differentiation capacities: 1) Biochemical assays, 2) Gene expression analysis, and 3) Morphological observations.Results: We found that both cells were able to differentiate into mature osteoblasts, as assayed by ALP activity. RT-PCR analysis showed the activation of the Opn gene after osteoblast differentiation. Morphological observations of both cells revealed the formation of black or dark-brown nodules after von Kossa staining. Nevertheless, only mononucleated cells showed the significant increase in TRAP activity characteristic of mature osteoclasts. The osteoclast-specific CatK gene was only upregulated in mononucleated cells. Morphological observations indicated the existence of multinucleated osteoclasts. Sca-1 was activated only in undifferentiated mononucleated cells, indicating that the cells were hematopoietic stem cells. In both cell lines, the housekeeping Gapdh gene was activated before and after differentiation.Conclusion: The isolated mononucleated cells were able to differentiate into both osteoblasts and osteoclasts; indicating that they are stem cells. On the other hand, MC3T3-E1 cells can only differentiate into osteoblasts; a characteristic of progenitor cells.",
author = "Yazid, {Muhammad D.} and {Zainal Ariffin}, {Shahrul Hisham} and Sahidan Senafi and Razak, {Mohamad A.} and {Megat Abdul Wahab}, Rohaya",
year = "2010",
month = "10",
day = "28",
doi = "10.1186/1475-2867-10-42",
language = "English",
volume = "10",
journal = "Cancer Cell International",
issn = "1475-2867",
publisher = "BioMed Central",

}

TY - JOUR

T1 - Determination of the differentiation capacities of murines' primary mononucleated cells and MC3T3-E1 cells

AU - Yazid, Muhammad D.

AU - Zainal Ariffin, Shahrul Hisham

AU - Senafi, Sahidan

AU - Razak, Mohamad A.

AU - Megat Abdul Wahab, Rohaya

PY - 2010/10/28

Y1 - 2010/10/28

N2 - Background: The main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus. The main purpose of this study was to determine the differentiation capacities of stem and progenitor cells. This study was performed using mononucleated cells originated from murine peripheral blood and MC3T3-E1 cells. Three approaches were used to determine their differentiation capacities: 1) Biochemical assays, 2) Gene expression analysis, and 3) Morphological observations.Results: We found that both cells were able to differentiate into mature osteoblasts, as assayed by ALP activity. RT-PCR analysis showed the activation of the Opn gene after osteoblast differentiation. Morphological observations of both cells revealed the formation of black or dark-brown nodules after von Kossa staining. Nevertheless, only mononucleated cells showed the significant increase in TRAP activity characteristic of mature osteoclasts. The osteoclast-specific CatK gene was only upregulated in mononucleated cells. Morphological observations indicated the existence of multinucleated osteoclasts. Sca-1 was activated only in undifferentiated mononucleated cells, indicating that the cells were hematopoietic stem cells. In both cell lines, the housekeeping Gapdh gene was activated before and after differentiation.Conclusion: The isolated mononucleated cells were able to differentiate into both osteoblasts and osteoclasts; indicating that they are stem cells. On the other hand, MC3T3-E1 cells can only differentiate into osteoblasts; a characteristic of progenitor cells.

AB - Background: The main morphological features of primitive cells, such as stem and progenitor cells, are that these cells consists only one nucleus. The main purpose of this study was to determine the differentiation capacities of stem and progenitor cells. This study was performed using mononucleated cells originated from murine peripheral blood and MC3T3-E1 cells. Three approaches were used to determine their differentiation capacities: 1) Biochemical assays, 2) Gene expression analysis, and 3) Morphological observations.Results: We found that both cells were able to differentiate into mature osteoblasts, as assayed by ALP activity. RT-PCR analysis showed the activation of the Opn gene after osteoblast differentiation. Morphological observations of both cells revealed the formation of black or dark-brown nodules after von Kossa staining. Nevertheless, only mononucleated cells showed the significant increase in TRAP activity characteristic of mature osteoclasts. The osteoclast-specific CatK gene was only upregulated in mononucleated cells. Morphological observations indicated the existence of multinucleated osteoclasts. Sca-1 was activated only in undifferentiated mononucleated cells, indicating that the cells were hematopoietic stem cells. In both cell lines, the housekeeping Gapdh gene was activated before and after differentiation.Conclusion: The isolated mononucleated cells were able to differentiate into both osteoblasts and osteoclasts; indicating that they are stem cells. On the other hand, MC3T3-E1 cells can only differentiate into osteoblasts; a characteristic of progenitor cells.

UR - http://www.scopus.com/inward/record.url?scp=77958537344&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77958537344&partnerID=8YFLogxK

U2 - 10.1186/1475-2867-10-42

DO - 10.1186/1475-2867-10-42

M3 - Article

VL - 10

JO - Cancer Cell International

JF - Cancer Cell International

SN - 1475-2867

M1 - 42

ER -