H19 gene is epigenetically stable in mouse multipotent germline stem cells

Area of Science:

• Epigenetics
• Stem Cell Biology
• Developmental Biology

Background:

• Testis-derived germline stem (GS) cells can be reprogrammed into multipotent GS (mGS) cells.
• mGS cells exhibit distinct DNA methylation patterns compared to GS cells.
• Understanding epigenetic alterations in mGS cells is vital for their application.

Purpose of the Study:

• To investigate the DNA methylation status of the H19 imprinting control region (ICR) in multipotent adult germline stem (maGS) cells.
• To determine how culture conditions affect epigenetic imprints in maGS cells.
• To assess the stability of epigenetic marks during in vitro culture and differentiation.

Main Methods:

• Established maGS cells were cultured with glial cell line-derived neurotrophic factor (GDNF) alone or with GDNF and leukemia inhibitory factor (LIF).
• DNA methylation analysis of the H19 ICR was performed using bisulfite sequencing PCR.
• mRNA transcript levels of Igf2 and H19 were quantified.

Main Results:

• The H19 ICR in maGS cells consistently showed hypermethylation with an androgenetic pattern, similar to GS cells.
• Igf2 mRNA levels were elevated, while H19 mRNA levels were reduced compared to embryonic stem cells.
• This androgenetic DNA methylation pattern remained stable through 54 passages and during differentiation via retinoic acid-induced embryoid bodies.

Conclusions:

• maGS cells exhibit epigenetic stability for the H19 gene under various in vitro culture and differentiation conditions.
• The observed epigenetic stability suggests potential for maGS cells in regenerative medicine.
• Further research into the epigenetic regulation and chromatin structure of maGS cells is warranted.