Gene expression patterns of Royan human embryonic stem cells correlate with their propensity and culture systems

Hassan Rassouli, Mona Khalaj, Seyedeh-Nafiseh Hassani, Shiva Nemati, Ghasem Hosseini Salekdeh, Hossein Baharvand

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Objective: Human embryonic stem cells (hESCs) have the potential to give rise to all types of cells in the human body when appropriately induced to differentiate. Stem cells can differentiate spontaneously into the three-germ layer derivatives by embryoid bodies (EBs) formation. However, the two-dimensional (2D) adherent culture of hESCs under defined conditions is commonly used for directed differentiation toward a specific type of mature cells. In this study, we aimed to determine the propensity of the Royan hESC lines based on comparison of expression levels of 46 lineage specific markers.

Materials and Methods: In this experimental study, we have compared the expression of lineage-specific markers in hESC lines during EB versus adherent-based spontaneous differentiation. We used quantitative real-time polymerase chain reaction (qRT-PCR) to assess expressions of 46 lineage-specific markers in 4 hESC lines, Royan H1 (RH1), RH2, RH5, and RH6, during spontaneous differentiation in both EB and adherent cultures at 0, 10, and 30 days after initiation of differentiation.

Results: Based on qRT-PCR data analysis, the liver and neuronal markers had higher expression levels in EBs, whereas skin-specific markers expressed at higher levels in the adherent culture. The results showed differential expression patterns of some lineage-specific markers in EBs compared with the adherent cultures.

Conclusion: According to these results, possibly the spontaneous differentiation technique could be a useful method for optimization of culture conditions to differentiate stem cells into specific cell types such ectoderm, neuron, endoderm and hepatocyte. This approach might prove beneficial for further work on maximizing the efficiency of directed differentiation and development of novel differentiation protocols.

LanguageEnglish
Pages290-299
Number of pages10
JournalCell Journal
Volume21
Issue number3
DOIs
Publication statusPublished - 2019

Fingerprint

Embryoid Bodies
Gene Expression
Cell Line
Real-Time Polymerase Chain Reaction
Stem Cells
Germ Layers
Endoderm
Ectoderm
Human Body
Hepatocytes
Human Embryonic Stem Cells
Neurons
Skin
Liver

Bibliographical note

Copyright the Author(s). Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • Differentiation
  • Gene Expression
  • Pluripotency
  • Propensity
  • Stem Cell

Cite this

Rassouli, Hassan ; Khalaj, Mona ; Hassani, Seyedeh-Nafiseh ; Nemati, Shiva ; Salekdeh, Ghasem Hosseini ; Baharvand, Hossein. / Gene expression patterns of Royan human embryonic stem cells correlate with their propensity and culture systems. In: Cell Journal. 2019 ; Vol. 21, No. 3. pp. 290-299.
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abstract = "Objective: Human embryonic stem cells (hESCs) have the potential to give rise to all types of cells in the human body when appropriately induced to differentiate. Stem cells can differentiate spontaneously into the three-germ layer derivatives by embryoid bodies (EBs) formation. However, the two-dimensional (2D) adherent culture of hESCs under defined conditions is commonly used for directed differentiation toward a specific type of mature cells. In this study, we aimed to determine the propensity of the Royan hESC lines based on comparison of expression levels of 46 lineage specific markers. Materials and Methods: In this experimental study, we have compared the expression of lineage-specific markers in hESC lines during EB versus adherent-based spontaneous differentiation. We used quantitative real-time polymerase chain reaction (qRT-PCR) to assess expressions of 46 lineage-specific markers in 4 hESC lines, Royan H1 (RH1), RH2, RH5, and RH6, during spontaneous differentiation in both EB and adherent cultures at 0, 10, and 30 days after initiation of differentiation. Results: Based on qRT-PCR data analysis, the liver and neuronal markers had higher expression levels in EBs, whereas skin-specific markers expressed at higher levels in the adherent culture. The results showed differential expression patterns of some lineage-specific markers in EBs compared with the adherent cultures. Conclusion: According to these results, possibly the spontaneous differentiation technique could be a useful method for optimization of culture conditions to differentiate stem cells into specific cell types such ectoderm, neuron, endoderm and hepatocyte. This approach might prove beneficial for further work on maximizing the efficiency of directed differentiation and development of novel differentiation protocols.",
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author = "Hassan Rassouli and Mona Khalaj and Seyedeh-Nafiseh Hassani and Shiva Nemati and Salekdeh, {Ghasem Hosseini} and Hossein Baharvand",
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Gene expression patterns of Royan human embryonic stem cells correlate with their propensity and culture systems. / Rassouli, Hassan; Khalaj, Mona; Hassani, Seyedeh-Nafiseh; Nemati, Shiva; Salekdeh, Ghasem Hosseini; Baharvand, Hossein.

In: Cell Journal, Vol. 21, No. 3, 2019, p. 290-299.

Research output: Contribution to journalArticleResearchpeer-review

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AU - Khalaj, Mona

AU - Hassani, Seyedeh-Nafiseh

AU - Nemati, Shiva

AU - Salekdeh, Ghasem Hosseini

AU - Baharvand, Hossein

N1 - Copyright the Author(s). Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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N2 - Objective: Human embryonic stem cells (hESCs) have the potential to give rise to all types of cells in the human body when appropriately induced to differentiate. Stem cells can differentiate spontaneously into the three-germ layer derivatives by embryoid bodies (EBs) formation. However, the two-dimensional (2D) adherent culture of hESCs under defined conditions is commonly used for directed differentiation toward a specific type of mature cells. In this study, we aimed to determine the propensity of the Royan hESC lines based on comparison of expression levels of 46 lineage specific markers. Materials and Methods: In this experimental study, we have compared the expression of lineage-specific markers in hESC lines during EB versus adherent-based spontaneous differentiation. We used quantitative real-time polymerase chain reaction (qRT-PCR) to assess expressions of 46 lineage-specific markers in 4 hESC lines, Royan H1 (RH1), RH2, RH5, and RH6, during spontaneous differentiation in both EB and adherent cultures at 0, 10, and 30 days after initiation of differentiation. Results: Based on qRT-PCR data analysis, the liver and neuronal markers had higher expression levels in EBs, whereas skin-specific markers expressed at higher levels in the adherent culture. The results showed differential expression patterns of some lineage-specific markers in EBs compared with the adherent cultures. Conclusion: According to these results, possibly the spontaneous differentiation technique could be a useful method for optimization of culture conditions to differentiate stem cells into specific cell types such ectoderm, neuron, endoderm and hepatocyte. This approach might prove beneficial for further work on maximizing the efficiency of directed differentiation and development of novel differentiation protocols.

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