Improved immunomagnetic enrichment of CD34(+) cells from umbilical cord blood using the CliniMACS cell separation system
| Autor: | Joseph Blake, Howard Voorhies, Daniel Weber, Ian B. Nicoud, Katherine A. Guthrie, Colleen Delaney, Shelly Heimfeld |
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| Rok vydání: | 2012 |
| Předmět: |
Cancer Research
Immunology CD34 Antigens CD34 Cell Count Cell Separation Biology Immunomagnetic separation Umbilical cord Article Flow cytometry Immunophenotyping Andrology Cell separation medicine Immunology and Allergy Humans Genetics (clinical) Transplantation medicine.diagnostic_test Immunomagnetic Separation Hematopoietic Stem Cell Transplantation Cell Biology Fetal Blood Flow Cytometry Billion Cells medicine.anatomical_structure Oncology Cord blood |
| Zdroj: | Cytotherapy. 14(7) |
| ISSN: | 1477-2566 |
| Popis: | Background aims CD34+ enrichment from cord blood units (CBU) is used increasingly in clinical applications involving ex vivo expansion. The CliniMACS instrument from Miltenyi Biotec is a current good manufacturing practice (cGMP) immunomagnetic selection system primarily designed for processing larger numbers of cells: a standard tubing set (TS) can process a maximum of 60 billion cells, while the larger capacity tubing set (LS) will handle 120 billion cells. In comparison, most CBU contain only 1–2 billion cells, raising a question regarding the optimal tubing set for CBU CD34+ enrichment. We compared CD34+ cell recovery and overall viability after CliniMACS processing of fresh CBU with either TS or LS. Methods Forty-six freshly collected CBU (≤ 36 h) were processed for CD34+ enrichment; 22 consecutive units were selected using TS and a subsequent 24 processed with LS. Cell counts and immunophenotyping were performed pre- and post-selection to assess total nucleated cells (TNC), viability and CD34+ cell content. Results Two-sample t -tests of mean CD34+ recovery and viability revealed significant differences in favor of LS (CD34+ recovery, LS = 56%, TS = 45%, P = 0.003; viability, LS = 74%, TS = 59%, P = 0.011). Stepwise linear regression, considering pre-processing unit age, viability, TNC and CD34+ purity, demonstrated statistically significant correlations only with the tubing set used and age of unit. Conclusions For CD34+ enrichment from fresh CBU, LS provided higher post-selection viability and more efficient recovery. In this case, a lower maximum TNC specification of TS was not predictive of better performance. The same may hold for smaller scale enrichment of other cell types with the CliniMACS instrument. |
| Databáze: | OpenAIRE |
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