Human Pluripotent and Progenitor Cells Display Cell Surface Cluster Differentiation Markers CD10, CD13, CD56, and MHC Class-I

Abstract

Each year millions of people suffer tissue loss or end-stage organ failure. While allogeneic therapies have saved and improved countless lives, they remain imperfect solutions. These therapies are limited by critical donor shortages, long-term morbidity, and mortality. A wide variety of transplants, congenital malformations, elective surgeries, and genetic disorders have the potential for treatment with autologous stem cells as a source of HLA-matched donor tissue. Our current research is aimed at characterizing cell surface cluster differentiation (CD) markers on human progenitor and pluripotent cells to aid in isolating comparatively purified populations of these cells. This study examined human pluripotent and progenitor cells isolated from fetal, mature, and geriatric individuals for the possible presence of 15 CD markers. The response to insulin and dexamethasone revealed that the cell isolates were composed of I i neage-com m itted progenitor cel Is and I i neage-u ncom m i tted pluri potent cells. Flow cytometry showed cell populations positive for CD10, CD13, CD56, and MHC Class-I markers and negative for CD3, CD5, CD7, CD11 b, CD14, CD15, CD16, CD19, CD25, CD45, and CD65 markers. Northern analysis revealed that CD13 and CD56 were actively transcribed at time of cell harvest. We report the first identification of CDlO, CD13, CD56, and MHC Class-l cell surface antigens on these human cells. [P.S.E.B.M. 1999[P.S.E.B.M. , Vol 2211 umerous studies have shown the existence of mesenchymal stem cells distributed widely N throughout the connective tissue compartments of many animals. These cells provide for the continued maintenance and repair of tissues throughout the life span of the individual. Examples of these cells include the unipotent

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

“…Conversely, stem cells derived from neural tissues (Bjornson et al, 1999) and muscle tissues (Jackson et al, 1999) can differentiate into blood. Furthermore, reserve stem cells derived from dermis can differentiate into skeletal muscle, fat, cartilage and bone (Young et al, 1999). These studies suggest that the fates of some of these reserve stem cells are not restricted by their location within a particular tissue or organ.…”

“…Cells derived from the connective tissues of skeletal muscle and dermis obtained from fetal and mature donors were purchased from Clonetics (San Diego, CA) and processed by differential plating and cryopreservation, yielding putative stem cells (Young et al, 1991(Young et al, , 1992a(Young et al, , 1999. In brief, the cells were transferred to plating medium-A (PM-A).…”

“…Geriatric cells were isolated from the endomysial, perimysial and epimysial connective tissue compartments associated with skeletal muscle specimens obtained from a 77-year-old female patient and a 67-year-old male patient following standard protocols for the isolation of mesenchymal stem cells Young et al, 1999). In brief, cells were liberated from the connective tissue compartment of skeletal muscle with collagenase (CLS-I, Worthington Biochemical Corp., Freehold, NJ) and dispase (Collaborative Biomedical Products, Bedford, MA).…”

“…We have previously characterized the capacity for differentiation of putative stem cells using an insulin/dexamethasone bioassay Pate et al, 1993;Rogers et al, 1995;Warejcka et al, 1996;Young, 2000;Young et al, 1991Young et al, , 1992aYoung et al, ,1992bYoung et al, , 1993Young et al, , 1995Young et al, , 1998aYoung et al, ,1998bYoung et al, , 1999. Numerous investigators have used antibodies to cluster of differentiation (CD) markers to characterize and isolate hematopoietic cells based on the profiles of their cell surface antigens (review, Kishimoto et al, 1997).…”

“…CD105 has been reported to be present on a tripotent adipogenicchondrogenic-osteogenic progenitor cell from the stroma of bone marrow (Barry et al, 1999;Pittenger et al, 1999). CD10, CD13, CD56, and MHC Class-I have been identified on human reserve stem cells (Young et al, 1999).…”