Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood

Current haematopoietic stem cell transplantation protocols rely heavily upon CD34+ cells to estimate haematopoietic stem and progenitor cell yield. However, several studies nowadays emerged with evidence that CD133+ cells represent a more primitive cell population than their CD34+ counterparts. Th...

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Main Authors:	Siti Norhaiza, H., Intan Zarina, Z.A., Rohaya, M.A.W., Sahidan, S., Shahrul Hisham, Z.A.
Format:	Article
Language:	English
Published:	Penerbit Universiti Kebangsaan Malaysia 2014
Online Access:	http://journalarticle.ukm.my/7297/ http://journalarticle.ukm.my/7297/ http://journalarticle.ukm.my/7297/1/43_1_14.pdf

id	ukm-7297
recordtype	eprints
spelling	ukm-72972016-12-14T06:43:40Z http://journalarticle.ukm.my/7297/ Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood Siti Norhaiza, H. Intan Zarina, Z.A. Rohaya, M.A.W. Sahidan, S. Shahrul Hisham, Z.A. Current haematopoietic stem cell transplantation protocols rely heavily upon CD34+ cells to estimate haematopoietic stem and progenitor cell yield. However, several studies nowadays emerged with evidence that CD133+ cells represent a more primitive cell population than their CD34+ counterparts. The objective of the present study was to isolate CD133+ stem cells by a rapid negative isolation method which depletes human peripheral blood mononuclear cells (MNCs) from cells expressing haematopoietic lineage markers CD2, CD3, CD11b, CD14,CD15, CD16, CD19, CD56, CD123, and CD235a and isolates a discrete lineage negative (Lin-) cell population (11±2% of MNC, n=6). Human peripheral blood MNCs were fractionated by density gradient centrifugation. After labelling with antibody against haematopoietic lineage markers, MNCs were subjected to magnetic assisted cell sorting (MACS). The expression of stem cells markers was examined by reverse transcriptase PCR (RT-PCR). We showed that Lin- stem-progenitor cell population encompassed the common markers used to characterize adult stem cells including SLAMF1, CD90 and CD133 and was also enriched for CD117, a surface receptor for stem cell factor. This cell population is more homogenous and primitive as compared to suspension MNCs which comprised of SLAMF1 and CD117 but the cells with haematopoietic lineage marker, CD14 and CD2 were also present. In conclusion, our approach was able to isolate a homogenous Lin– stem-progenitor cell population from human peripheral blood hence offers a promising alternative to current haematopoietic stem and progenitor selection methods. Penerbit Universiti Kebangsaan Malaysia 2014-06 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/7297/1/43_1_14.pdf Siti Norhaiza, H. and Intan Zarina, Z.A. and Rohaya, M.A.W. and Sahidan, S. and Shahrul Hisham, Z.A. (2014) Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood. Malaysian Applied Biology, 43 (1). pp. 125-132. ISSN 0126-8643 http://mabjournal.com/
repository_type	Digital Repository
institution_category	Local University
institution	Universiti Kebangasaan Malaysia
building	UKM Institutional Repository
collection	Online Access
language	English
description	Current haematopoietic stem cell transplantation protocols rely heavily upon CD34+ cells to estimate haematopoietic stem and progenitor cell yield. However, several studies nowadays emerged with evidence that CD133+ cells represent a more primitive cell population than their CD34+ counterparts. The objective of the present study was to isolate CD133+ stem cells by a rapid negative isolation method which depletes human peripheral blood mononuclear cells (MNCs) from cells expressing haematopoietic lineage markers CD2, CD3, CD11b, CD14,CD15, CD16, CD19, CD56, CD123, and CD235a and isolates a discrete lineage negative (Lin-) cell population (11±2% of MNC, n=6). Human peripheral blood MNCs were fractionated by density gradient centrifugation. After labelling with antibody against haematopoietic lineage markers, MNCs were subjected to magnetic assisted cell sorting (MACS). The expression of stem cells markers was examined by reverse transcriptase PCR (RT-PCR). We showed that Lin- stem-progenitor cell population encompassed the common markers used to characterize adult stem cells including SLAMF1, CD90 and CD133 and was also enriched for CD117, a surface receptor for stem cell factor. This cell population is more homogenous and primitive as compared to suspension MNCs which comprised of SLAMF1 and CD117 but the cells with haematopoietic lineage marker, CD14 and CD2 were also present. In conclusion, our approach was able to isolate a homogenous Lin– stem-progenitor cell population from human peripheral blood hence offers a promising alternative to current haematopoietic stem and progenitor selection methods.
format	Article
author	Siti Norhaiza, H. Intan Zarina, Z.A. Rohaya, M.A.W. Sahidan, S. Shahrul Hisham, Z.A.
spellingShingle	Siti Norhaiza, H. Intan Zarina, Z.A. Rohaya, M.A.W. Sahidan, S. Shahrul Hisham, Z.A. Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
author_facet	Siti Norhaiza, H. Intan Zarina, Z.A. Rohaya, M.A.W. Sahidan, S. Shahrul Hisham, Z.A.
author_sort	Siti Norhaiza, H.
title	Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
title_short	Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
title_full	Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
title_fullStr	Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
title_full_unstemmed	Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
title_sort	molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood
publisher	Penerbit Universiti Kebangsaan Malaysia
publishDate	2014
url	http://journalarticle.ukm.my/7297/ http://journalarticle.ukm.my/7297/ http://journalarticle.ukm.my/7297/1/43_1_14.pdf
first_indexed	2023-09-18T19:49:19Z
last_indexed	2023-09-18T19:49:19Z
_version_	1777406112249675776

Molecular characterization of a lineage negative stem-progenitor cell population from human peripheral blood

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