Hyaluronan as a novel marker for rapid selection of connective tissue progenitors.
Connective Tissue Progenitors (CTPs) in bone and marrow tissue are an important therapeutic target for bone regeneration. Concentration and selection of CTPs from the heterogenous population of cells in bone marrow remains a challenge due to their low prevalence. This study identifies surface-bound hyaluronan (HA), a component of the in vivo niche for CTPs, and evaluates HA as a useful surface marker for positive selection of CTPs. Mononuclear cells from bone marrow were labeled and magnetically separated on the basis of hyaluronan binding. HA(+++), HA(+) and HA(-) fractions were cultured and assayed for colony formation using a quantitative image-processing system. A mean of 2.7% of cells were retained in the HA(+++) fraction and were enriched by 3.4-fold (range of 95% CI: 2.3-4.8) in CTP prevalence when compared to the unselected buffy-coated bone marrow aspirate (BCM). In addition, colonies formed by HA(+++) CTPs demonstrated greater proliferation (more cells per colony) and greater alkaline phosphatase activity than CTP colonies derived from unselected BCM. These data demonstrate that one or more subsets of human marrow-derived CTPs retain a HA rich matrix on their surface at the time of harvest. HA(+++) CTPs may offer a useful cell population for regenerative therapies.
Department of Biomedical Engineering, Orthopaedic and Rheumatological Research Center, Lerner Research Institute, The Cleveland Clinic, 9620 Carnegie, Cleveland, OH 44195, USA. carallt@ccf.org
Tonya
T
Caralla
Cynthia
C
Boehm
Vincent
V
Hascall
George
G
Muschler
D058621:Blood Buffy Coat / Q000166:cytology* / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D003239:Connective Tissue Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism; D005260:Female; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry*; D008297:Male; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
D006820:Hyaluronic Acid