In vitro expansion of fetal liver hematopoietic stem cells

Abstract Fetal liver hematopoietic stem and progenitor cells (HSPCs) have been considered appropriate for the management of aplastic anemia owing to their proliferative potential. Bone marrow recovery was possible in some cases; the engraftment potential of these cells, however was unsatisfactory, p...

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Autores principales:	Rashmi Bhardwaj, Lalit Kumar, Deepika Chhabra, N. K. Mehra, Atul sharma, Sujata Mohanty, Vinod Kochupillai
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b0b024f5e2464d888cbfb70b2086d52f
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spelling	oai:doaj.org-article:b0b024f5e2464d888cbfb70b2086d52f2021-12-02T17:50:41ZIn vitro expansion of fetal liver hematopoietic stem cells10.1038/s41598-021-91272-62045-2322https://doaj.org/article/b0b024f5e2464d888cbfb70b2086d52f2021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91272-6https://doaj.org/toc/2045-2322Abstract Fetal liver hematopoietic stem and progenitor cells (HSPCs) have been considered appropriate for the management of aplastic anemia owing to their proliferative potential. Bone marrow recovery was possible in some cases; the engraftment potential of these cells, however was unsatisfactory, possibly due to the availability of a smaller number of these cells from a single fetus. The present study explores how we can expand fetal liver hematopoietic stem cells under in vitro conditions. We isolated mononuclear cells from fetal liver and hematopoietic stem cells were identified and analyzed by cell surface marker CD34. CD34+ fetal liver HSPCs cells were separated by magnetic cell sorting positive selection method. HSPCs (CD34+) were cultured by using 5 cytokines, stem cell factor (SCF), granulocyte macrophages-colony stimulating factor (GM-CSF), interleukin-6 (IL-6), Fms-related tyrosine kinase 3 (FLT-3) and erythropoietin (EPO), in 4 different combinations along with supplements, in serum-free culture media for 21 days. Cell viability continued to be greater than 90% throughout 21 days of culture. The cells expanded best in a combination of media, supplements and 5 cytokines, namely SCF, FLT-3, IL6, EPO and GM-CSF to yield a large number of total (CD34+ & CD34-) cells. Even though the total number of nucleated cells increased in culture significantly, levels of CD34 antigen expression declined steadily over this period.Rashmi BhardwajLalit KumarDeepika ChhabraN. K. MehraAtul sharmaSujata MohantyVinod KochupillaiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rashmi Bhardwaj Lalit Kumar Deepika Chhabra N. K. Mehra Atul sharma Sujata Mohanty Vinod Kochupillai In vitro expansion of fetal liver hematopoietic stem cells
description	Abstract Fetal liver hematopoietic stem and progenitor cells (HSPCs) have been considered appropriate for the management of aplastic anemia owing to their proliferative potential. Bone marrow recovery was possible in some cases; the engraftment potential of these cells, however was unsatisfactory, possibly due to the availability of a smaller number of these cells from a single fetus. The present study explores how we can expand fetal liver hematopoietic stem cells under in vitro conditions. We isolated mononuclear cells from fetal liver and hematopoietic stem cells were identified and analyzed by cell surface marker CD34. CD34+ fetal liver HSPCs cells were separated by magnetic cell sorting positive selection method. HSPCs (CD34+) were cultured by using 5 cytokines, stem cell factor (SCF), granulocyte macrophages-colony stimulating factor (GM-CSF), interleukin-6 (IL-6), Fms-related tyrosine kinase 3 (FLT-3) and erythropoietin (EPO), in 4 different combinations along with supplements, in serum-free culture media for 21 days. Cell viability continued to be greater than 90% throughout 21 days of culture. The cells expanded best in a combination of media, supplements and 5 cytokines, namely SCF, FLT-3, IL6, EPO and GM-CSF to yield a large number of total (CD34+ & CD34-) cells. Even though the total number of nucleated cells increased in culture significantly, levels of CD34 antigen expression declined steadily over this period.
format	article
author	Rashmi Bhardwaj Lalit Kumar Deepika Chhabra N. K. Mehra Atul sharma Sujata Mohanty Vinod Kochupillai
author_facet	Rashmi Bhardwaj Lalit Kumar Deepika Chhabra N. K. Mehra Atul sharma Sujata Mohanty Vinod Kochupillai
author_sort	Rashmi Bhardwaj
title	In vitro expansion of fetal liver hematopoietic stem cells
title_short	In vitro expansion of fetal liver hematopoietic stem cells
title_full	In vitro expansion of fetal liver hematopoietic stem cells
title_fullStr	In vitro expansion of fetal liver hematopoietic stem cells
title_full_unstemmed	In vitro expansion of fetal liver hematopoietic stem cells
title_sort	in vitro expansion of fetal liver hematopoietic stem cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/b0b024f5e2464d888cbfb70b2086d52f
work_keys_str_mv	AT rashmibhardwaj invitroexpansionoffetalliverhematopoieticstemcells AT lalitkumar invitroexpansionoffetalliverhematopoieticstemcells AT deepikachhabra invitroexpansionoffetalliverhematopoieticstemcells AT nkmehra invitroexpansionoffetalliverhematopoieticstemcells AT atulsharma invitroexpansionoffetalliverhematopoieticstemcells AT sujatamohanty invitroexpansionoffetalliverhematopoieticstemcells AT vinodkochupillai invitroexpansionoffetalliverhematopoieticstemcells
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In vitro expansion of fetal liver hematopoietic stem cells

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