Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin.
Using novel media formulations, it has been demonstrated that human placenta and umbilical cord blood-derived CD34+ cells can be expanded and differentiated into erythroid cells with high efficiency. However, obtaining mature and functional erythrocytes from the immature cell cultures with high puri...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3428333?pdf=render |
| _version_ | 1818421773665828864 |
|---|---|
| author | Xiaoxia Jin Stewart Abbot Xiaokui Zhang Lin Kang Vanessa Voskinarian-Berse Rui Zhao Marina V Kameneva Lee R Moore Jeffrey J Chalmers Maciej Zborowski |
| author_facet | Xiaoxia Jin Stewart Abbot Xiaokui Zhang Lin Kang Vanessa Voskinarian-Berse Rui Zhao Marina V Kameneva Lee R Moore Jeffrey J Chalmers Maciej Zborowski |
| author_sort | Xiaoxia Jin |
| collection | DOAJ |
| description | Using novel media formulations, it has been demonstrated that human placenta and umbilical cord blood-derived CD34+ cells can be expanded and differentiated into erythroid cells with high efficiency. However, obtaining mature and functional erythrocytes from the immature cell cultures with high purity and in an efficient manner remains a significant challenge. A distinguishing feature of a reticulocyte and maturing erythrocyte is the increasing concentration of hemoglobin and decreasing cell volume that results in increased cell magnetophoretic mobility (MM) when exposed to high magnetic fields and gradients, under anoxic conditions. Taking advantage of these initial observations, we studied a noninvasive (label-free) magnetic separation and analysis process to enrich and identify cultured functional erythrocytes. In addition to the magnetic cell separation and cell motion analysis in the magnetic field, the cell cultures were characterized for cell sedimentation rate, cell volume distributions using differential interference microscopy, immunophenotyping (glycophorin A), hemoglobin concentration and shear-induced deformability (elongation index, EI, by ektacytometry) to test for mature erythrocyte attributes. A commercial, packed column high-gradient magnetic separator (HGMS) was used for magnetic separation. The magnetically enriched fraction comprised 80% of the maturing cells (predominantly reticulocytes) that showed near 70% overlap of EI with the reference cord blood-derived RBC and over 50% overlap with the adult donor RBCs. The results demonstrate feasibility of label-free magnetic enrichment of erythrocyte fraction of CD34+ progenitor-derived cultures based on the presence of paramagnetic hemoglobin in the maturing erythrocytes. |
| first_indexed | 2024-12-14T13:15:41Z |
| format | Article |
| id | doaj.art-b91322a913ee475dbb5495a7d96428b1 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T13:15:41Z |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-b91322a913ee475dbb5495a7d96428b12022-12-21T23:00:04ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0178e3949110.1371/journal.pone.0039491Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin.Xiaoxia JinStewart AbbotXiaokui ZhangLin KangVanessa Voskinarian-BerseRui ZhaoMarina V KamenevaLee R MooreJeffrey J ChalmersMaciej ZborowskiUsing novel media formulations, it has been demonstrated that human placenta and umbilical cord blood-derived CD34+ cells can be expanded and differentiated into erythroid cells with high efficiency. However, obtaining mature and functional erythrocytes from the immature cell cultures with high purity and in an efficient manner remains a significant challenge. A distinguishing feature of a reticulocyte and maturing erythrocyte is the increasing concentration of hemoglobin and decreasing cell volume that results in increased cell magnetophoretic mobility (MM) when exposed to high magnetic fields and gradients, under anoxic conditions. Taking advantage of these initial observations, we studied a noninvasive (label-free) magnetic separation and analysis process to enrich and identify cultured functional erythrocytes. In addition to the magnetic cell separation and cell motion analysis in the magnetic field, the cell cultures were characterized for cell sedimentation rate, cell volume distributions using differential interference microscopy, immunophenotyping (glycophorin A), hemoglobin concentration and shear-induced deformability (elongation index, EI, by ektacytometry) to test for mature erythrocyte attributes. A commercial, packed column high-gradient magnetic separator (HGMS) was used for magnetic separation. The magnetically enriched fraction comprised 80% of the maturing cells (predominantly reticulocytes) that showed near 70% overlap of EI with the reference cord blood-derived RBC and over 50% overlap with the adult donor RBCs. The results demonstrate feasibility of label-free magnetic enrichment of erythrocyte fraction of CD34+ progenitor-derived cultures based on the presence of paramagnetic hemoglobin in the maturing erythrocytes.http://europepmc.org/articles/PMC3428333?pdf=render |
| spellingShingle | Xiaoxia Jin Stewart Abbot Xiaokui Zhang Lin Kang Vanessa Voskinarian-Berse Rui Zhao Marina V Kameneva Lee R Moore Jeffrey J Chalmers Maciej Zborowski Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin. PLoS ONE |
| title | Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin. |
| title_full | Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin. |
| title_fullStr | Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin. |
| title_full_unstemmed | Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin. |
| title_short | Erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin. |
| title_sort | erythrocyte enrichment in hematopoietic progenitor cell cultures based on magnetic susceptibility of the hemoglobin |
| url | http://europepmc.org/articles/PMC3428333?pdf=render |
| work_keys_str_mv | AT xiaoxiajin erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT stewartabbot erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT xiaokuizhang erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT linkang erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT vanessavoskinarianberse erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT ruizhao erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT marinavkameneva erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT leermoore erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT jeffreyjchalmers erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin AT maciejzborowski erythrocyteenrichmentinhematopoieticprogenitorcellculturesbasedonmagneticsusceptibilityofthehemoglobin |