title: Molecular and phenotypic characterization of CD133 and SSEA4 enriched very small embryonic-like stem cells in human cord blood.
abstract: Very small embryonic-like stem cells (VSELs) are immature primitive cells residing in adult and fetal tissues. This study describes enrichment strategy and molecular and phenotypic characterization of human cord blood VSELs. Flow cytometry analysis revealed that a majority of VSELs (LIN(-)/CD45(-)/CD34(+)) were present in the red blood cell (RBC) pellet after Ficoll-Hypaque centrifugation in contrast to the hematopoietic stem cells (LIN(-)/CD45(+)/CD34(+)) in the interphase layer. Thus, after lyses of RBCs, VSELs were enriched using CD133 and SSEA4 antibodies. These enriched cells were small in size (4-6 μm), spherical, exhibited telomerase activity and expressed pluripotent stem cell (OCT4A, OCT4, SSEA4, NANOG, SOX2, REX1), primordial germ cell (STELLA, FRAGILIS) as well as primitive hematopoietic (CD133, CD34) markers at protein and transcript levels. Heterogeneity was noted among VSELs based on subtle differences in expression of various markers studied. DNA analysis and cell cycle studies revealed that a majority of enriched VSELs were diploid, non-apoptotic and in G0/G1 phase, reflecting their quiescent state. VSELs also survived 5-fluorouracil treatment in vitro and treated cells entered into cell cycle. This study provides further support for the existence of pluripotent, diploid and relatively quiescent VSELs in cord blood and suggests further exploration of the subpopulations among them.
authors.affiliation: Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India.
authors.forename: A
authors.initials: A
authors.lastname: Shaikh
authors.affiliation: Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India.
authors.forename: P
authors.initials: P
authors.lastname: Nagvenkar
authors.affiliation: Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India.
authors.forename: P
authors.initials: P
authors.lastname: Pethe
authors.affiliation: Jaslok Hospital & Research Centre, Mumbai, India.
authors.forename: I
authors.initials: I
authors.lastname: Hinduja
authors.affiliation: Stem Cell Biology Department, National Institute for Research in Reproductive Health, Mumbai, India.
authors.forename: D
authors.initials: D
authors.lastname: Bhartiya
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism*; D000954:Antigens, Surface / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002453:Cell Cycle; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D005312:Fetal Blood / Q000166:cytology*; D005472:Fluorouracil / Q000494:pharmacology; D006023:Glycoproteins / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D016130:Immunophenotyping; D010455:Peptides / Q000378:metabolism*; D010641:Phenotype; D011003:Ploidies; D055746:Stage-Specific Embryonic Antigens / Q000378:metabolism*
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D000954:Antigens, Surface; D015415:Biomarkers; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; D055746:Stage-Specific Embryonic Antigens; C066584:stage-specific embryonic antigen-4; D005472:Fluorouracil