Lectin microarray analysis of pluripotent and multipotent stem cells

Abstract

Stem cells have a capability to self-renew and differentiate into multiple types of cells; specific markers are available to identify particular stem cells for developmental biology research. In this study, we aimed to define the status of somatic stem cells and the pluripotency of human embryonic stem (hES) and induced pluripotent stem (iPS) cells using a novel molecular methodology, lectin microarray analysis. Our lectin microarray analysis successfully categorized murine somatic stem cells into the appropriate groups of differentiation potency. We then classified hES and iPS cells by the same approach. Undifferentiated hES cells were clearly distinguished from differentiated hES cells after embryoid formation. The pair-wise comparison means based on 'false discovery rate' revealed that three lectins -Euonymus europaeus lectin (EEL), Maackia amurensis lectin (MAL) and Phaseolus vulgaris leucoagglutinin [PHA(L)]-generated maximal values to define undifferentiated and differentiated hES cells. Furthermore, to define a pluripotent stem cell state, we generated a discriminant for the undifferentiated state with pluripotency. The discriminant function based on lectin reactivities was highly accurate for judgment of stem cell pluripotency. These results suggest that glycomic analysis of stem cells leads to a novel comprehensive approach for quality control in cell-based therapy and regenerative medicine.

The effects of vincristine sulfate on expression of galectin-3, Bcl-2, and carbohydrate structures specific for EEL, GSL-1, and RCA-1 lectins in bitches with naturally occurring canine transmissible venereal tumor

“…Different biological functions in animals are attributed to lectins, including regulation of cell adhesion and glycoprotein synthesis. Pluripotent stem cells had the specific glycan structure recognized by EEL multipotent cell types (28). GSL-I has been reported to bind several glycoproteins, including laminin, and is necessary for intercellular communication (29).…”

Podocalyxin-Targeting Comparative Glycan Profiling Reveals Difference between Human Embryonic Stem Cells and Embryonal Carcinoma Cells

“…Although the theoretical molecular mass of podocalyxin is merely 55 kDa, actually immunoprecipitated podocalyxin was found to be >200 kDa, probably with heavily glycosylation consistent with previous reports that the molecular mass of podocalyxin is extremely large representing a feature of sialomucin [5,7]. In our previous works using lectin microarray, we successfully categorized murine and human somatic stem cells into groups based on differentiation potencies [43,44]. Considering potential roles of glycans in cell-cell interactions, it is effective to distinguish hESCs from hECCs based on the glycan profiles targeting a major and heavily glycosylated glycoprotein, i.e., podocalyxin.…”

“…In the authors' laboratory, as a lectin-based glycan profiling method, a unique lectin microarray system based on an evanescentfield activated fluorescence detection principle was developed [41,42]. With this system, we and other groups successfully profiled distinct sets of glycomes of both somatic and pluripotent stem cells [43][44][45][46][47].…”

A practical guide to induced pluripotent stem cell research using patient samples

“…In addition to these standard quality controls, profiling of RNA expression, DNA methylation and glycans can be added for monitoring when necessary. 10,13,36,37 These comprehensive analyses would also elucidate pathogenic states such as aberrant genomic methylation and gene expression of patient iPSCs.…”