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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 CD34+/CD45+ cells from UCB, PBMNC and BM, after isolation (Day 0) and following 5 days of expansion.The Q2 gate represents the population double positive for CD34 (APC) and CD45 (PE).
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pone-0112496-g001: FACS analyses of the CD34+/CD45+ cells from UCB, PBMNC and BM, after isolation (Day 0) and following 5 days of expansion.The Q2 gate represents the population double positive for CD34 (APC) and CD45 (PE).

Mentions: We next tested if larger numbers of reticulocytes could be obtained from ex vivo expanded CD34+-enriched cell populations. After 5 days of expansion in serum-free medium with TPO and SCF, the total cell populations increased >10-fold in cultures initiated with UCB/CD34+-enriched cells, 3-fold for BM/CD34+-enriched cells while for PBMC no expansion was observed (Table 1, 3 independent experiments). FACS analysis demonstrated an increase in the CD34+/CD45+ population between Day 0 and Day 5 for all three cell sources: from 55% to 70% (SD±2) for UCB, 35% to 55% (SD±5) for BM, and 16% to 29% (SD±16) for PBMC (n = 3 for CB and BM, n = 2 for PBMC)(Figure 1).


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 CD34+/CD45+ cells from UCB, PBMNC and BM, after isolation (Day 0) and following 5 days of expansion.The Q2 gate represents the population double positive for CD34 (APC) and CD45 (PE).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0112496-g001: FACS analyses of the CD34+/CD45+ cells from UCB, PBMNC and BM, after isolation (Day 0) and following 5 days of expansion.The Q2 gate represents the population double positive for CD34 (APC) and CD45 (PE).
Mentions: We next tested if larger numbers of reticulocytes could be obtained from ex vivo expanded CD34+-enriched cell populations. After 5 days of expansion in serum-free medium with TPO and SCF, the total cell populations increased >10-fold in cultures initiated with UCB/CD34+-enriched cells, 3-fold for BM/CD34+-enriched cells while for PBMC no expansion was observed (Table 1, 3 independent experiments). FACS analysis demonstrated an increase in the CD34+/CD45+ population between Day 0 and Day 5 for all three cell sources: from 55% to 70% (SD±2) for UCB, 35% to 55% (SD±5) for BM, and 16% to 29% (SD±16) for PBMC (n = 3 for CB and BM, n = 2 for PBMC)(Figure 1).

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