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Evaluation of Reliable Reference Genes for In Vitro Erythrocyte Generation from Cord Blood CD34+ Cells.
Xu, Lei; Gao, Zhan; Yang, Zhou; Qu, Mingyi; Li, Huilin; Chen, Lin; Lv, Yang; Fan, Zeng; Yue, Wen; Li, Cuiying; Xie, Xiaoyan; Pei, Xuetao.
Afiliación
  • Xu L; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing, China.
  • Gao Z; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou, China.
  • Yang Z; Clinical Medical College of Air Force, Anhui Medical University, Hefei, China.
  • Qu M; Air Force Medical Center, PLA, Beijing, China.
  • Li H; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing, China.
  • Chen L; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou, China.
  • Lv Y; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing, China.
  • Fan Z; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou, China.
  • Yue W; Beijing Institute of Radiation Medicine, Beijing, China.
  • Li C; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing, China.
  • Xie X; South China Research Center for Stem Cell & Regenerative Medicine, SCIB, Guangzhou, China.
  • Pei X; Stem Cell and Regenerative Medicine Lab, Institute of Health Service and Transfusion Medicine, Beijing, China.
DNA Cell Biol ; 40(9): 1200-1210, 2021 Sep.
Article en En | MEDLINE | ID: mdl-34227876
In vitro generation of red blood cells has the potential to circumvent shortfalls in the global demand for blood for transfusion applications. However, cell differentiation and proliferation are often regulated by precise changes in gene expression, but the underlying mechanisms and molecular changes remain unclear. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) can be used to evaluate multiple target genes. To make the results more reliable, suitable reference genes should be used to calibrate the error associated with qRT-PCR. In this study, we utilized bioinformatics to screen 3 novel candidate reference genes (calcium and integrin binding family member 2 [CIB2], olfactory receptor family 8 subfamily B member 8 [OR8B8], and zinc finger protein 425 [ZNF425]) along with eight traditional reference genes (glyceraldehyde-3-phosphate dehydrogenase [GAPDH], ß-actin [ACTB], 18S RNA, ß2-microglobulin [ß2-MG], peptidylprolyl isomerase A [PPIA], TATA box-binding protein [TBP], hydroxymethylbilane synthase [HMBS], and hypoxanthine phosphoribosyltransferase 1 [HPRT1]). Two software algorithms (geNorm and NormFinder) were used to evaluate the stability of expression of the 11 genes at different stages of erythrocyte development. Comprehensive analysis showed that expression of GAPDH and TBP was the most stable, whereas ZNF425 and OR8B8 were the least suitable candidate genes. These results suggest that appropriate reference genes should be selected before performing gene expression analysis during erythroid differentiation and that GAPDH and TBP are suitable reference genes for gene expression studies on erythropoiesis.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Represoras / Células Madre / Proteínas de Unión al Calcio / Eritrocitos Límite: Humans Idioma: En Revista: DNA Cell Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2021 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Proteínas Represoras / Células Madre / Proteínas de Unión al Calcio / Eritrocitos Límite: Humans Idioma: En Revista: DNA Cell Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2021 Tipo del documento: Article País de afiliación: China Pais de publicación: Estados Unidos