Separation of human bone marrow by counterflow centrifugation monitored by DNA‐flowcytometry

Abstract

Human bone marrow was fractionated by counterflow centrifugation into 16 fractions with increasing cell size. Three distinct subpopulations could be recognized: small lymphocytic cells, medium-sized nucleated erythroid cells and large myeloid elements. DNA-flowcytometry and 3H-thymidine uptake showed that within the erythroid and myeloid cell populations counterflow centrifugation separates each population according to the cell cycle phase. Hypotonic treatment of bone marrow for removal of the erythroid nucleated cells resulted in a complete abrogation of the proliferating erythroid cell population. Counterflow centrifugation also separates the small non-proliferating myeloid and erythroid committed stem cells from the larger proliferating stem cells. It appeared feasible to separate the small lymphocytic cells from the majority of BFU-E and CFU-GM, due to the larger size of the proliferating normoblasts and the committed progenitor cells. Elimination of the mature lymphocytes from the haematopoietic stem cells by counterflow centrifugation may offer an alternative approach to the prevention of graft versus host disease (GvHD).

Ca2+ and caspases are involved in hydroxyl radical-induced apoptosis in erythrocytes of Jian carp (Cyprinus carpio var. Jian)

“…In the present study, carp erythrocytes from the peripheral blood were separated into three fractions using the fixed-angle centrifugation. The cells in different developmental stages can be identified by measuring the cell cycle using the flow cytometer (Witte et al 1984). The cycle of eukaryotic cells is divided into G 1 , S, G 2 and M phases (Cooper 2000).…”

Identification of self-renewing myoblasts in the progeny of single human muscle satellite cells

“…Native HMSC, which are in the G0 phase of the cell cycle, are known to be small cells [1,17]. Moreover, physical features such as different size and light scatter properties distinguish hematopoietic or keratinocytes stem cells from differentiated cells [8,9,12,28]. Finally, a characteristic of stem cells, as seen in many different types of renewing cell populations, is that under steady-state conditions, they are non-dividing [25].…”