Characterization of multipotent neural progenitors isolated from human adult gut.

<p>(A) Bright-field view of representative adult enteric neurosphere-like bodies (NLBs) after 14 days <i>in vitro</i>. Free-floating NLBs could be observed with an estimated average number of 35 NLBs per preparation and a sphere diameter of 71.6 μm. Different NLB phenotypes include round- (star, comparable to B), ellipsoid- (white arrow, comparable to C) or irregular-shaped (black arrow, comparable to D) NLBs. (B–D) Depending to the individual NLB phenotype, different cellular compositions can be derived as shown for the ENS marker gene p75 in immunocytochemistry. (E–G) Proliferative NLB cells also incorporated BrdU and expressed previously described progenitor markers including Sox2 and Nestin. (H–J) After 2 weeks of differentiation NLBs built a monolayer composed of neuronal (TuJ), glial (S100β) and smooth muscle-like cells (SMA). (K–M) Molecular cytogenetic FISH analyses revealed no chromosomal alterations as representatively shown for three chromosomal interphase nuclei. (N) A schematic illustration of a chromosome is shown. In total, 15 different probe sets recognizing 41 different unique subtelomeric repeats were applied. (O) Efficacy and cellular distribution of NLBs strongly depends to the individual preparation, relying on age, amount and quality of tissue, as demonstrated by qPCR analysis for three representative marker genes: p75, Ret and tropomyosin (TPM). Scale bars represent in (A) 100 μm, (B–G) 25 μm, (H–J) 50 μm.