Your browser doesn't support javascript.
loading
Improving hematopoietic differentiation from human induced pluripotent stem cells by the modulation of Hippo signaling with a diarylheptanoid derivative.
Thongsa-Ad, Umnuaychoke; Wongpan, Anongnat; Wongkummool, Wasinee; Chaiwijit, Phaewa; Uppakara, Kwanchanok; Chaiyakitpattana, Gorawin; Singpant, Passanan; Tong-Ngam, Pirut; Chukhan, Amnat; Pabuprappap, Wachirachai; Wongniam, Sirapope; Suksamrarn, Apichart; Hongeng, Suradej; Anurathapan, Usanarat; Kulkeaw, Kasem; Tubsuwan, Alisa; Bhukhai, Kanit.
Afiliación
  • Thongsa-Ad U; Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
  • Wongpan A; Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
  • Wongkummool W; Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.
  • Chaiwijit P; Center of Multidisciplinary Technology for Advanced Medicine (CMUTEAM), Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand.
  • Uppakara K; Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
  • Chaiyakitpattana G; Chakri Naruebodindra Medical Institute, Faculty of Medicine Ramathibodi Hospital, Mahidol University, Samut Prakan, 10540, Thailand.
  • Singpant P; Department of Physiology, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
  • Tong-Ngam P; Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.
  • Chukhan A; Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.
  • Pabuprappap W; Prima Scientific, 147/170-171 Baromrajchonnee, Arunamarin, Bangkok, 10700, Thailand.
  • Wongniam S; Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, 10240, Thailand.
  • Suksamrarn A; Center for Scientific Instrumentation and Platform Services Unit, Faculty of Science, Mahidol University, Bangkok, 10400, Thailand.
  • Hongeng S; Department of Chemistry and Center of Excellence for Innovation in Chemistry, Faculty of Science, Ramkhamhaeng University, Bangkok, 10240, Thailand.
  • Anurathapan U; Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, 10400, Thailand.
  • Kulkeaw K; Department of Pediatrics, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, 10400, Thailand.
  • Tubsuwan A; Siriraj Integrative Center for Neglected Parasitic Diseases, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, 10700, Thailand.
  • Bhukhai K; Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom, 73170, Thailand.
Stem Cell Res Ther ; 15(1): 60, 2024 Mar 03.
Article en En | MEDLINE | ID: mdl-38433217
ABSTRACT

BACKGROUND:

The diarylheptanoid ASPP 049 has improved the quality of adult hematopoietic stem cell (HSC) expansion ex vivo through long-term reconstitution in animal models. However, its effect on hematopoietic regeneration from human induced pluripotent stem cells (hiPSCs) is unknown.

METHOD:

We utilized a defined cocktail of cytokines without serum or feeder followed by the supplementation of ASPP 049 to produce hematopoietic stem/progenitor cells (HSPCs). Flow cytometry and trypan blue exclusion analysis were used to identify nonadherent and adherent cells. Nonadherent cells were harvested to investigate the effect of ASPP 049 on multipotency using LTC-IC and CFU assays. Subsequently, the mechanism of action was explored through transcriptomic profiles, which were validated by qRT-PCR, immunoblotting, and immunofluorescence analysis.

RESULT:

The supplementation of ASPP 049 increased the number of phenotypically defined primitive HSPCs (CD34+CD45+CD90+) two-fold relative to seeded hiPSC colonies, indicating enhanced HSC derivation from hiPSCs. Under ASPP 049-supplemented conditions, we observed elevated HSPC niches, including CD144+CD73- hemogenic- and CD144+CD73+ vascular-endothelial progenitors, during HSC differentiation. Moreover, harvested ASPP 049-treated cells exhibited improved self-renewal and a significantly larger proportion of different blood cell colonies with unbiased lineages, indicating enhanced HSC stemness properties. Transcriptomics and KEGG analysis of sorted CD34+CD45+ cells-related mRNA profiles revealed that the Hippo signaling pathway is the most significant in responding to WWTR1/TAZ, which correlates with the validation of the protein expression. Interestingly, ASPP 049-supplemented HSPCs upregulated 11 genes similarly to umbilical cord blood-derived HSPCs.

CONCLUSION:

These findings suggest that ASPP 049 can improve HSC-generating protocols with proliferative potentials, self-renewal ability, unbiased differentiation, and a definable mechanism of action for the clinical perspective of hematopoietic regenerative medicine.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Madre Pluripotentes Inducidas / Vía de Señalización Hippo Límite: Adult / Animals / Humans Idioma: En Revista: Stem Cell Res Ther Año: 2024 Tipo del documento: Article País de afiliación: Tailandia Pais de publicación: Reino Unido
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Células Madre Pluripotentes Inducidas / Vía de Señalización Hippo Límite: Adult / Animals / Humans Idioma: En Revista: Stem Cell Res Ther Año: 2024 Tipo del documento: Article País de afiliación: Tailandia Pais de publicación: Reino Unido