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Hematopoietic stem/progenitor cell sources to generate reticulocytes for Plasmodium vivax culture.

Noulin F, Manesia JK, Rosanas-Urgell A, Erhart A, Borlon C, Van Den Abbeele J, d'Alessandro U, Verfaillie CM - PLoS ONE (2014)

Bottom Line: We here compared the production of reticulocytes using an established in vitro culture system from three different sources of hematopoietic stem/progenitor cells (HSPC), i.e. umbilical cord blood (UCB), bone marrow (BM) and adult peripheral blood (PB).Compared to CD34+-enriched populations of PB and BM, CD34+-enriched populations of UCB produced the highest amount of reticulocytes that could be invaded by P. vivax.As invasion by P. vivax was significantly better in reticulocytes generated in vitro, we also suggest that P. vivax may have a preference for invading immature reticulocytes, which should be confirmed in future studies.

View Article: PubMed Central - PubMed

Affiliation: Unit of Malariology, Institute of Tropical Medicine, Antwerp, Belgium.

ABSTRACT
The predilection of Plasmodium vivax (P. vivax) for reticulocytes is a major obstacle for its establishment in a long-term culture system, as this requires a continuous supply of large quantities of reticulocytes, representing only 1-2% of circulating red blood cells. We here compared the production of reticulocytes using an established in vitro culture system from three different sources of hematopoietic stem/progenitor cells (HSPC), i.e. umbilical cord blood (UCB), bone marrow (BM) and adult peripheral blood (PB). Compared to CD34+-enriched populations of PB and BM, CD34+-enriched populations of UCB produced the highest amount of reticulocytes that could be invaded by P. vivax. In addition, when CD34+-enriched cells were first expanded, a further extensive increase in reticulocytes was seen for UCB, to a lesser degree BM but not PB. As invasion by P. vivax was significantly better in reticulocytes generated in vitro, we also suggest that P. vivax may have a preference for invading immature reticulocytes, which should be confirmed in future studies.

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FACS analyses of the CD235a+/CD71+ cells from UCB, PBMNC and BM, after 5 days of expansion and 14 days of differentiation.The Q2 gate represents the population positive for CD235a (Per-CP-Cy5-5) and CD71 (PE).
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pone-0112496-g002: FACS analyses of the CD235a+/CD71+ cells from UCB, PBMNC and BM, after 5 days of expansion and 14 days of differentiation.The Q2 gate represents the population positive for CD235a (Per-CP-Cy5-5) and CD71 (PE).

Mentions: Following expansion, a similar number of cells (irrespective of the CD34+ content or expansion) were cultured under reticulocyte differentiation conditions. After 7 days of erythroid differentiation, the total number of cells, previously subject to an expansion step, was 3 times higher compared to CD34+ cells that were immediately induced to differentiate. After 14 days of differentiation, expanded cells expressed high levels of CD235a and CD71 receptors, regardless of cell source (respectively 87.4% for UCB, 81.7% for BM and 70.6% for PB; Figure 2). Compared to unexpanded cells, the proportion of reticulocytes obtained at day 14 from in vitro expanded CD34+ cells was 5 to 10-fold higher.


Hematopoietic stem/progenitor cell sources to generate reticulocytes for Plasmodium vivax culture.

Noulin F, Manesia JK, Rosanas-Urgell A, Erhart A, Borlon C, Van Den Abbeele J, d'Alessandro U, Verfaillie CM - PLoS ONE (2014)

FACS analyses of the CD235a+/CD71+ cells from UCB, PBMNC and BM, after 5 days of expansion and 14 days of differentiation.The Q2 gate represents the population positive for CD235a (Per-CP-Cy5-5) and CD71 (PE).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4231068&req=5

pone-0112496-g002: FACS analyses of the CD235a+/CD71+ cells from UCB, PBMNC and BM, after 5 days of expansion and 14 days of differentiation.The Q2 gate represents the population positive for CD235a (Per-CP-Cy5-5) and CD71 (PE).
Mentions: Following expansion, a similar number of cells (irrespective of the CD34+ content or expansion) were cultured under reticulocyte differentiation conditions. After 7 days of erythroid differentiation, the total number of cells, previously subject to an expansion step, was 3 times higher compared to CD34+ cells that were immediately induced to differentiate. After 14 days of differentiation, expanded cells expressed high levels of CD235a and CD71 receptors, regardless of cell source (respectively 87.4% for UCB, 81.7% for BM and 70.6% for PB; Figure 2). Compared to unexpanded cells, the proportion of reticulocytes obtained at day 14 from in vitro expanded CD34+ cells was 5 to 10-fold higher.

Bottom Line: We here compared the production of reticulocytes using an established in vitro culture system from three different sources of hematopoietic stem/progenitor cells (HSPC), i.e. umbilical cord blood (UCB), bone marrow (BM) and adult peripheral blood (PB).Compared to CD34+-enriched populations of PB and BM, CD34+-enriched populations of UCB produced the highest amount of reticulocytes that could be invaded by P. vivax.As invasion by P. vivax was significantly better in reticulocytes generated in vitro, we also suggest that P. vivax may have a preference for invading immature reticulocytes, which should be confirmed in future studies.

View Article: PubMed Central - PubMed

Affiliation: Unit of Malariology, Institute of Tropical Medicine, Antwerp, Belgium.

ABSTRACT
The predilection of Plasmodium vivax (P. vivax) for reticulocytes is a major obstacle for its establishment in a long-term culture system, as this requires a continuous supply of large quantities of reticulocytes, representing only 1-2% of circulating red blood cells. We here compared the production of reticulocytes using an established in vitro culture system from three different sources of hematopoietic stem/progenitor cells (HSPC), i.e. umbilical cord blood (UCB), bone marrow (BM) and adult peripheral blood (PB). Compared to CD34+-enriched populations of PB and BM, CD34+-enriched populations of UCB produced the highest amount of reticulocytes that could be invaded by P. vivax. In addition, when CD34+-enriched cells were first expanded, a further extensive increase in reticulocytes was seen for UCB, to a lesser degree BM but not PB. As invasion by P. vivax was significantly better in reticulocytes generated in vitro, we also suggest that P. vivax may have a preference for invading immature reticulocytes, which should be confirmed in future studies.

Show MeSH
Related in: MedlinePlus