Evaluation of a Soybean Lectin–based Method for the Enrichment of Erythroblasts

Abstract

S U M M A R YWe performed a comparative study of the enrichment of erythroblasts by a soybean agglutinin galactose-specific lectin method and a standardized magnetic cell-sorting (MACS) protocol. Blood samples, obtained from 11 pregnant women at between 11 and 40 weeks of gestation, were split and examined by each method in parallel. The number of erythroblasts recovered by the lectin method was approximately eightfold higher than the number obtained by MACS. Our data suggest that the lectin-based method may provide a better approach for the enrichment of rare fetal erythroblasts from maternal blood.

A microfluidics approach for the isolation of nucleated red blood cells (NRBCs) from the peripheral blood of pregnant women

“…Among the 37 pregnancies with unknown follow-up, a mean of 37.68 NRBC/mL, median of 21.08, and a range of 0.37 to 168.2 was identified. These results were compared with those of previously reported studies using alternative enrichment processes (Table 2) (Slunga-Tallberg et al ., 1995; Takabayashi et al ., 1995; Babochkina et al ., 2005; Purwosunu et al ., 2006b; Kwon et al ., 2007; Mavrou et al ., 2007; Sekizawa et al ., 2007). Mean NRBC/mL in those studies ranged from 0.40 to 5.40 vs 37.68 in the current study.…”

“…This is in contrast with means ranging from 0.58–5.4 NRBC/mL identified by MACS density gradient, and in two of three studies using the galactose-lectin method. In one study, as many as 2.00 to 59.00 NRBCs were identified using soybean agglutinin galactose-lectin (Babochkina et al ., 2005). However, the current method revealed an approximate 10- to 20-fold increase in NRBC counts over any other previously reported method.…”

“…The most frequently used methods to enrich or sort NRBCs from the whole blood of pregnant women have included density gradient separation (Kuo and Guo, 1999; Sekizawa et al ., 1999; Samura et al ., 2000; Smits et al ., 2000), fluorescence-activated cell sorting (FACS)-based positive selection (Iverson et al ., 1981; Bianchi et al ., 1990; Price et al ., 1991; Lewis et al ., 1996; Wang et al ., 2000), magnetic-activated cell sorting (MACS)-based depletion or enrichment (Ganshirt-Ahlert et al ., 1992, 1993; Busch et al ., 1994; Campagnoli et al ., 1997; Valerio et al ., 1997; Troeger et al ., 1999b; Mavrou et al ., 2007), micromanipulation (Takabayashi et al ., 1995; Sekizawa et al ., 2007), selective lysis (de Graaf et al ., 1999), and galactose-lectin-based methods (Babochkina et al ., 2005; Purwosunu et al ., 2006b; Sekizawa et al ., 2007). Both FACS and MACS rely on sequential steps of preliminary separation of background cells, generally by density gradient centrifugation or cell lysis, followed by antigen–antibody recognition using NRBCspecific monoclonal antibodies.…”

Current and emerging techniques of fetal cell separation from maternal blood

“…The highest number of NRBCs recovered from maternal blood was achieved by Wachtel et al in 1996 and again in 1998 using charge flow separation (CFS) [63,64]. In the first case over 2000 fetal NRBCs were recovered from 20 ml samples of maternal blood.…”

Situación actual del diagnóstico prenatal no invasivo

“…Por último, se ha descrito la utilización de una lectina que se une específicamente a los residuos de galactosa presentes en la superficie celular del eritroblasto 46 .…”