Analysis of Stem Cell Engraftment Markers
To determine the utility of pluripotent stem cells for cell transplantation, it is critical to detect stem cell engraftment and to track their differentiation into specialized cells. The ability to genetically manipulate stem cells to express the reporter gene Green Fluorescent Protein [GFP], coupled with immunological detection of GFP, permits such analysis (Min et al., 2002, J. Appl. Physiol; Tang et al., 2002, Cell. Mol. Neurobiol.; Orlic et al., 2001, Ann. NY Acad. Sci.; Hess et al., 2002, Stroke; Lee et al., 2001, Mol. Ther; Englund, 2002, Exp. Neurol). This technique was recently used to demonstrate the use of human ES cells to improve ventricular function in infarcted hearts (Min et al., 2002, J. Appl. Physiol) and to differentiate into astrocytes (Tang et al., 2002, Cell. Mol. Neurobiol). Additionally, this technique has been used to demonstrate the ability of bone marrow stem cells to repair myocardial (Orlic et al., 2001, Ann. NY Acad. Sci) and cerebral Infarction (Hess et al., 2002, Stroke) to monitor mesenchymal stem cell differentiation following transplantation (Lee et al., 2001, Mol. Ther) and to monitor the in vivo differentiation of transplanted neural precursor cells (Englund et al., 2002, Exp. Neurol).
Engrafted neurons derived from human NSCs can be discriminated from host tissue using Human- specific antibodies to human N-CAM (Millipore cat. no. MAB2122), human mitochondria (Cat. No. MAB1273) or human nuclei (Millipore cat. no. MAB1281) (Uchida et al., 2000, PNAS). Detection of the antigen Ki-67, a nuclear protein expressed by cells in all phases of the active cycle and absent in the resting phase, allows for the determination of proliferating cells of human origin (Uchida et al., 2000, PNAS). The detection of implanted, pre-differentiated cells can be accomplished by pre-labeling proliferating and differentiating cells with BrdU prior to implantation (Vescovi et al., 1999, Exp. Neurol). Resulting neurons and glia can be detected using an anti-BrdU antibody.
Millipore’s products provide researchers with a range of options for the detection of engrafted stem cells.
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