Open AccessHydrogen sulfide facilitates reprogramming and trans-differentiation in 3D dermal fibroblast
Author(s) -
Elena A. Ostrakhovitch,
Shin Akakura,
Siamak Tabibzadeh
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0241685
Subject(s) - reprogramming , homeobox protein nanog , microbiology and biotechnology , sox2 , gene knockdown , lin28 , klf4 , chemistry , cellular differentiation , biology , cell culture , fibroblast , cell , induced pluripotent stem cell , biochemistry , embryonic stem cell , gene , genetics
The efficiency of cell reprogramming in two-dimensional (2D) cultures is limited. Given that cellular stemness is intimately related to microenvironmental changes, 3D cell cultures have the potential of overcoming this limited capacity by allowing cells to self-organize by aggregation. In 3D space, cells interact more efficiently, modify their cellular topology, gene expression, signaling, and metabolism. It is yet not clear as how 3D culture environments modify the reprogramming potential of fibroblasts. We demonstrate that 3D spheroids from dermal fibroblasts formed under ultra-low attachment conditions showed increased lactate production. This is a requisite for cell reprogramming, increase their expression of pluripotency genes, such as OCT4 , NANOG and SOX2 , and display upregulated cystathionine-β-synthase ( CBS ) and hydrogen sulfide (H 2 S) production. Knockdown of CBS by RNAi suppresses lactic acid and H 2 S production and concomitantly decreases the expression of OCT4 and NANOG . On the contrary, H 2 S donors, NaHS and garlic-derived diallyl trisulfide (DATS), promote the expression of OCT4 , and support osteogenic trans-differentiation of fibroblasts. These results demonstrate that CBS mediated release of H 2 S regulates the reprogramming of dermal fibroblasts grown in 3D cultures and supports their trans-differentiation.