title: Current and historical perspectives on methodological flaws in processing umbilical cord blood.
abstract: Umbilical cord blood (UCB) hematopoietic stem cells (HSC-CD34+) are valuable for treating malignant or nonmalignant disease. Processing UCB by HESPAN-6% and anti-CD34-Miltenyi particles provides insufficient cells for treating adults. Physicochemical-electrokinetic studies on UCB-mononuclear cells (MNCs) under conditions of delayed processing, ice or very low temperatures, and some cell separation media identified artifacts introduced by procedures. Adsorption of biomaterials from cell damage by temperature, degradation products after using enzymes, harsh reagents, dithiothreitol, and HESPAN affect cell properties and distribution. Miltenyi particles internalized by cells could release iron that accumulating in liver or spleen would then risk toxicity. Summary topics included the effects of temperature, HESPAN (fast sedimenting agent), glycoproteases, DNase, and dithiothreitol risk affecting cell receptors in recognition, "homing," leading to possible unintended iatrogenic bioeffects should such cells be transfused into humans. The loss of undetectable and uncaptured low CD34 antigen-bearing cells by Miltenyi particles seems to occur when the current methods of isolation of CD34+ cells and other cells are critically assessed. The purpose here is to highlight and suggest avoiding the procedural flaws involved. Preventing ice temperatures avoids ice-damaged platelets releasing biomaterials that are adsorbed on cells altering UBC-MNCs/HSC properties and cell loss. Omitting the positive selection with antibody-linked Miltenyi particles obviates the use of harsh reagents to release the cells. Internalized Miltenyi particles are a toxicity hazard that needs investigations. Achieving approximately 5% yields of CD34+ cells (153 × 10(5) /110 mL cord-placenta blood) is a major advance holding great promise, for the first time increasing the prospect of stem cell therapy of 70-kg adults, using a single UCB donation (with dose of 1.5 × 10(5) cells/kg) and considerably cheaper cultured red blood cells manufacture (multiple packs/2 × 10(12) ).
authors: University of Cambridge, Cambridge, UK.
authors: J N
authors: JN
authors: Mehrishi
mesh_terms: D000327:Adsorption; D000328:Adult; D018952:Antigens, CD34 / Q000032:analysis; D001800:Blood Specimen Collection; D002469:Cell Separation / Q000379:methods*; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D006239:Haplotypes; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006650:Histocompatibility Testing; D006801:Humans; D006895:Hydroxyethyl Starch Derivatives; D011247:Pregnancy
chemical_list: D018952:Antigens, CD34; D006895:Hydroxyethyl Starch Derivatives