Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis
Erythropoiesis is a highly regulated process and undergoes several genotypic and phenotypic changes during differentiation. The phenotypic changes can be evaluated using a combination of cell surface markers expressed at different cellular stages of erythropoiesis using FACS. However, limited studie...
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MDPI AG
2023-05-01
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| author | Shobhita Katiyar Arunim Shah Khaliqur Rahman Naresh Kumar Tripathy Rajesh Kashyap Soniya Nityanand Chandra Prakash Chaturvedi |
| author_facet | Shobhita Katiyar Arunim Shah Khaliqur Rahman Naresh Kumar Tripathy Rajesh Kashyap Soniya Nityanand Chandra Prakash Chaturvedi |
| author_sort | Shobhita Katiyar |
| collection | DOAJ |
| description | Erythropoiesis is a highly regulated process and undergoes several genotypic and phenotypic changes during differentiation. The phenotypic changes can be evaluated using a combination of cell surface markers expressed at different cellular stages of erythropoiesis using FACS. However, limited studies are available on the in-depth phenotypic characterization of progenitors from human adult hematopoietic stem and progenitor cells (HSPCs) to red blood cells. Therefore, using a set of designed marker panels, in the current study we have kinetically characterized the hematopoietic, erythroid progenitors, and terminally differentiated erythroblasts ex vivo. Furthermore, the progenitor stages were explored for expression of CD117, CD31, CD41a, CD133, and CD45, along with known key markers CD36, CD71, CD105, and GPA. Additionally, we used these marker panels to study the stage-specific phenotypic changes regulated by the epigenetic regulator; <i>Nuclear receptor binding SET Domain protein 1</i> (<i>NSD1</i>) during erythropoiesis and to study ineffective erythropoiesis in myelodysplastic syndrome (MDS) and pure red cell aplasia (PRCA) patients. Our immunophenotyping strategy can be used to sort and study erythroid-primed hematopoietic and erythroid precursors at specified time points and to study diseases resulting from erythroid dyspoiesis. Overall, the current study explores the in-depth kinetics of phenotypic changes occurring during human erythropoiesis and applies this strategy to study normal and defective erythropoiesis. |
| first_indexed | 2024-03-11T04:21:55Z |
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| id | doaj.art-28e0aac856e0403b88ee604fc204601d |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-11T04:21:55Z |
| publishDate | 2023-05-01 |
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| series | Cells |
| spelling | doaj.art-28e0aac856e0403b88ee604fc204601d2023-11-17T22:44:03ZengMDPI AGCells2073-44092023-05-01129130310.3390/cells12091303Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective ErythropoiesisShobhita Katiyar0Arunim Shah1Khaliqur Rahman2Naresh Kumar Tripathy3Rajesh Kashyap4Soniya Nityanand5Chandra Prakash Chaturvedi6Stem Cell Research Center, Department of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaStem Cell Research Center, Department of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaDepartment of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaDepartment of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaDepartment of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaDepartment of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaStem Cell Research Center, Department of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Lucknow 226014, IndiaErythropoiesis is a highly regulated process and undergoes several genotypic and phenotypic changes during differentiation. The phenotypic changes can be evaluated using a combination of cell surface markers expressed at different cellular stages of erythropoiesis using FACS. However, limited studies are available on the in-depth phenotypic characterization of progenitors from human adult hematopoietic stem and progenitor cells (HSPCs) to red blood cells. Therefore, using a set of designed marker panels, in the current study we have kinetically characterized the hematopoietic, erythroid progenitors, and terminally differentiated erythroblasts ex vivo. Furthermore, the progenitor stages were explored for expression of CD117, CD31, CD41a, CD133, and CD45, along with known key markers CD36, CD71, CD105, and GPA. Additionally, we used these marker panels to study the stage-specific phenotypic changes regulated by the epigenetic regulator; <i>Nuclear receptor binding SET Domain protein 1</i> (<i>NSD1</i>) during erythropoiesis and to study ineffective erythropoiesis in myelodysplastic syndrome (MDS) and pure red cell aplasia (PRCA) patients. Our immunophenotyping strategy can be used to sort and study erythroid-primed hematopoietic and erythroid precursors at specified time points and to study diseases resulting from erythroid dyspoiesis. Overall, the current study explores the in-depth kinetics of phenotypic changes occurring during human erythropoiesis and applies this strategy to study normal and defective erythropoiesis.https://www.mdpi.com/2073-4409/12/9/1303erythropoiesisflowcytometrymarker panels |
| spellingShingle | Shobhita Katiyar Arunim Shah Khaliqur Rahman Naresh Kumar Tripathy Rajesh Kashyap Soniya Nityanand Chandra Prakash Chaturvedi Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis Cells erythropoiesis flowcytometry marker panels |
| title | Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis |
| title_full | Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis |
| title_fullStr | Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis |
| title_full_unstemmed | Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis |
| title_short | Analysis of Immunophenotypic Changes during Ex Vivo Human Erythropoiesis and Its Application in the Study of Normal and Defective Erythropoiesis |
| title_sort | analysis of immunophenotypic changes during ex vivo human erythropoiesis and its application in the study of normal and defective erythropoiesis |
| topic | erythropoiesis flowcytometry marker panels |
| url | https://www.mdpi.com/2073-4409/12/9/1303 |
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