Human reserve pluripotent mesenchymal stem cells are present in the connective tissues of skeletal muscle and dermis derived from fetal, adult, and geriatric donors

Abstract

This study details the profile of 13 cell surface cluster differentiation markers on human reserve stem cells derived from connective tissues. Stem cells were isolated from the connective tissues of dermis and skeletal muscle derived from fetal, mature, and geriatric humans. An insulin/dexamethasone phenotypic bioassay was used to determine the identity of the stem cells from each population. All populations contained lineage-committed myogenic, adipogenic, chondrogenic, and osteogenic progenitor stem cells as well as lineageuncommitted pluripotent stem cells capable of forming muscle, adipocytes, cartilage, bone, fibroblasts, and endothelial cells. Flow cytometric analysis of adult stem cell populations revealed positive staining for CD34 and CD90 and negative staining for CD3, CD4, CD8, CD11c, CD33, CD36, CD38, CD45, CD117, Glycophorin-A, and HLA DR-II. Anat Rec 264: [51][52][53][54][55][56][57][58][59][60][61][62] 2001.

Clonogenic analysis reveals reserve stem cells in postnatal mammals. II. Pluripotent epiblastic‐like stem cells

“…Actual doubling time during the exponential postconfluent growth phase approximated 12-14 hr. This was in contrast to an 18-to 24-hr exponential preconfluent growth phase for either pluripotent mesenchymal stem cells or germ layer lineage mesodermal stem cells that become contact inhibited at confluence (Young et al, 2001a(Young et al, , 2001b. Cells were aliquoted at 10 6 -10 7 cells/ml and cryopreserved.…”

“…The identity of specific types of progenitor and pluripotent cells within an unknown population of cells can be ascertained by comparing the effects of treatment with a progression factor and a general nonspecific lineage-induction agent (Young et al, 1992a(Young et al, , 1992b(Young et al, , 1998a(Young et al, , 1998b(Young et al, , 2001a(Young et al, , 2001bLucas et al, 1993Lucas et al, , 1995Pate et al, 1993;Rogers et al, 1995;Warejcka et al, 1996;Young, 2000Young and Black, 2004). Progression factors, such as insulin (at 2-5 g/ml), accelerate phenotypic expression in progenitor cells but have no effect on the induction of phenotypic expression in pluripotent stem cells.…”

“…Cell types belonging to embryonic, ectodermal, mesodermal, and endodermal lineages were assayed using previously established morphological, histochemical, and immunochemical procedures to denote changes in phenotypic expression markers (Young et al, 1991(Young et al, , 1992a(Young et al, , 1992b(Young et al, , 1998a(Young et al, , 1998b(Young et al, , 2001a(Young et al, , 2001bYoung, 2000Young and Black, 2004) (Table 1).…”

“…Scl-40␤ clone was plated into gelatinized 96-well plates at 10 3 cells per well in CM and allowed to attach for 24 hr (Young et al, 2001a(Young et al, , 2001b. CM consisted of 89% (v/v) Opti-MEM-based medium (catalog no.…”

“…27 White fat, also denoted as unilocular adipose tissue, was identified as a mononucleated cell with a peripherally-located nucleus and containing a large central intracellular vacuole filled with refractile lipid and stained histochemically for saturated neutral lipid using Oil Red-O (Sigma) and Sudan Black-B (Chroma-Gesellschaft, Roboz Surgical Co., Washington, DC) (Young et al, 2001a). 28 Brown fat, also denoted as multi-locular adipose tissue, was identified as a mononucleated cell with a centrally-located nucleus containing multiple small intracellular vacuoles filled with refractile lipid and stained histochemically for saturated neutral lipid using Oil Red-O (Sigma) and Sudan Black-B (Chroma-Gesellschaft) (Young, 2000;Young et al, 2001b). 29 -33 Cartilage: structures thought to be cartilage nodules were tentatively identified as aggregates of rounded cells containing pericellular matrix halos.…”