CD133 immunomagnetic separation: effectiveness of the method for CD133(+) isolation from umbilical cord blood.

Publisher: Elsevier Country of Publication: England NLM ID: 100895309 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1477-2566 (Electronic) Linking ISSN: 14653249 NLM ISO Abbreviation: Cytotherapy Subsets: MEDLINE

Publication: 2013- : London : Elsevier
Original Publication: Oxford, England : ISIS Medical Media, c1999-

Antigens, CD/*metabolism ; Fetal Blood/*cytology ; Glycoproteins/*metabolism ; Immunomagnetic Separation/*methods ; Peptides/*metabolism; AC133 Antigen ; Animals ; Antigens, CD34/metabolism ; Cell Separation ; Flow Cytometry ; Leukocytes, Mononuclear/cytology ; Mice ; Microspheres

Background Aims: Umbilical cord blood (UCB) is a rich source of stem cells, the characterization and isolation of which requires specific stem cell markers and reliable and reproducible protocols.
Methods: We assessed CD133 isolation in 39 UCB samples, using a commercial immunomagnetic cell-sorting protocol, and, because of its non-reproducibility, we applied optimized protocols in an effort to improve it. These included extra-labeling of the selected CD133(+) subpopulation and indirect labeling using anti-phycoerythrin (PE) microbeads, goat anti-mouse IgG microbeads or a combination of both. The CD34 isolation was used as a control.
Results: The mononuclear cell fraction expressed 0.53±0.06% CD133. The corresponding value for CD34 was 1.64±0.15%. Following the manufacturer's instructions, the CD34 isolation resulted in a population expressing 93±1.25% CD34 while, after the corresponding process, CD133(+) expression ranged from 10% to 85% (median 60%). The optimized isolation protocols did not result in improved CD133(+) yield. The variation in the purity of the CD133 population cannot be attributed to the different clones of CD133 used, because they do not cross-block, while other factors such as glycosylation, which could possibly interfere, do not apply in normal hematopoietic stem cells (HSC).
Conclusions: CD34 isolation by the immunomagnetic method results in highly pure CD34(+) population, while the efficient and reproducible yield of a pure CD133(+) population is not feasible. Therefore quantification of the positive cells should follow each isolation procedure in order to confirm the number of CD133(+) cells.

0 (AC133 Antigen)
0 (Antigens, CD)
0 (Antigens, CD34)
0 (Glycoproteins)
0 (Peptides)
0 (Prom1 protein, mouse)

Date Created: 20120315 Date Completed: 20121101 Latest Revision: 20180629

20250114

10.3109/14653249.2012.663487

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