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Elevated expression of DNMT1 is associated with increased expansion and proliferation of hematopoietic stem cells co-cultured with human MSCs
Blood Research ; : 25-30, 2017.
Article in English | WPRIM | ID: wpr-226886
ABSTRACT

BACKGROUND:

Mesenchymal stem cells (MSCs) play an important role in hematopoietic stem cell (HSC) maintenance, proliferation, and apoptosis. DNA methyltransferase 1 (DNMT1) is considered an essential factor in the maintenance of HSCs in mammalian cells. Therefore, this study was conducted to evaluate the mRNA expression level of DNMT1 during cord blood (CB)-HSC ex vivo expansion with MSCs.

METHODS:

Ex vivo cultures of CB-HSCs were performed in three culture conditions for 7 days cytokines, cytokines with MSCs, and only MSCs. Total and viable cell numbers were counted after 5 and 7 days using trypan blue stain, and the stem cell percentage was then evaluated by flow cytometry. Moreover, in vitro colony-forming unit assay was carried out to detect clonogenic potential of HSCs at days 0 and 7 using MethoCult H4434. Finally, DNMT1 mRNA expression level was evaluated by real-time polymerase chain reaction.

RESULTS:

Maximum CB-CD34⁺ cell expansion was observed on day 7 in all the three cultures. After 7 days, ex vivo expansion of CB-CD34⁺ cells indicated a significant decrease in DNMT1 expression in the cytokine cultures, whereas in the two co-culture conditions DNMT1 expression was increased. A significant difference between the number of CD34⁺ and CD34⁻ cells in the cytokine co-culture system was observed.

CONCLUSION:

These data indicated that an elevated expression of DNMT1 is associated with increased expansion and proliferation of HSCs co-cultured with human MSCs. Hence, DNMT1 may be a potential factor in the maintenance of expanded HSCs co-cultured with human MSCs.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Stem Cells / Trypan Blue / In Vitro Techniques / DNA / RNA, Messenger / Hematopoietic Stem Cells / Cell Count / Cytokines / Apoptosis / Coculture Techniques Limits: Humans Language: English Journal: Blood Research Year: 2017 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Stem Cells / Trypan Blue / In Vitro Techniques / DNA / RNA, Messenger / Hematopoietic Stem Cells / Cell Count / Cytokines / Apoptosis / Coculture Techniques Limits: Humans Language: English Journal: Blood Research Year: 2017 Type: Article