In vitro proliferation of mononucleated suspension and adherent cells from mouse and human peripheral blood system

Shahrul Hisham Zainal Ariffin, Muhammad Dain Yazid, Ruzanna Ab Kadir, Shabnam Kermani, Rohaya Megat Abdul Wahab

Research output: Contribution to journalArticle

Abstract

Primary cells have a limited proliferative capacity with a finite number of times as compared with cell line which can grow indefinitely. Therefore, this study was carried out to identify the proliferative capacity of primary mononucleated cells from mouse and human. The mononucleated cells were isolated from mouse and human peripheral blood by density gradient centrifugation using Ficoll-Paque Plus. The two types of cells i.e. suspension and adherent forms were obtained after culturing the isolated mononucleated cells for 4 days in the complete medium consists of Alpha Minimal Essential Medium, 10% newborn calf serum and 2% penicillin/streptomycin. The cells were then cultured for another 10 days to observe cell viability using trypan blue exclusion assay (suspension form) and MTT assay (adherent form). NSO and MC3T3-E1 cell lines were selected as control cell for suspension and adherent cells, respectively. Our results showed that the proliferation rate of mouse suspension mononucleated cells increased from 1.31 ± 0.24 cells/day (day 5) to 2.69 ± 0.42 cells/day (day 10) whilst, for human suspension cells, the proliferation rate slightly increased from 0.56 ± 0.20 cells/day (day 5) to 0.76 ± 0.29 cells/day (day 10). However, the proliferation rate of mouse adherent mononucleated cells decreased from 0.23 ± 0.02 cells/day (day 5) to 0.17 ± 0.01 cells/day (day 10). Meanwhile, human adherent cells maintained proliferation rate at approximately 0.67 ± 0.18 cells/day. In conclusion, adherent primary mononucleated cells from both mouse and human have limited capacity to generate more cells in vitro as compared with suspension mononucleated cells.

Original languageEnglish
Pages (from-to)1099-1107
Number of pages9
JournalSains Malaysiana
Volume41
Issue number9
Publication statusPublished - Sep 2012

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Suspensions
In Vitro Techniques
Cell Proliferation
Cell Line
Ficoll
Trypan Blue
Density Gradient Centrifugation
Streptomycin
Penicillins
Cell Survival

Keywords

  • Adherent cells
  • Peripheral blood
  • Proliferation
  • Suspension cells

ASJC Scopus subject areas

  • General

Cite this

In vitro proliferation of mononucleated suspension and adherent cells from mouse and human peripheral blood system. / Zainal Ariffin, Shahrul Hisham; Yazid, Muhammad Dain; Kadir, Ruzanna Ab; Kermani, Shabnam; Megat Abdul Wahab, Rohaya.

In: Sains Malaysiana, Vol. 41, No. 9, 09.2012, p. 1099-1107.

Research output: Contribution to journalArticle

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AB - Primary cells have a limited proliferative capacity with a finite number of times as compared with cell line which can grow indefinitely. Therefore, this study was carried out to identify the proliferative capacity of primary mononucleated cells from mouse and human. The mononucleated cells were isolated from mouse and human peripheral blood by density gradient centrifugation using Ficoll-Paque Plus. The two types of cells i.e. suspension and adherent forms were obtained after culturing the isolated mononucleated cells for 4 days in the complete medium consists of Alpha Minimal Essential Medium, 10% newborn calf serum and 2% penicillin/streptomycin. The cells were then cultured for another 10 days to observe cell viability using trypan blue exclusion assay (suspension form) and MTT assay (adherent form). NSO and MC3T3-E1 cell lines were selected as control cell for suspension and adherent cells, respectively. Our results showed that the proliferation rate of mouse suspension mononucleated cells increased from 1.31 ± 0.24 cells/day (day 5) to 2.69 ± 0.42 cells/day (day 10) whilst, for human suspension cells, the proliferation rate slightly increased from 0.56 ± 0.20 cells/day (day 5) to 0.76 ± 0.29 cells/day (day 10). However, the proliferation rate of mouse adherent mononucleated cells decreased from 0.23 ± 0.02 cells/day (day 5) to 0.17 ± 0.01 cells/day (day 10). Meanwhile, human adherent cells maintained proliferation rate at approximately 0.67 ± 0.18 cells/day. In conclusion, adherent primary mononucleated cells from both mouse and human have limited capacity to generate more cells in vitro as compared with suspension mononucleated cells.

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