An adaptation of a previously reported flow cytometric technique is described. This technique was applied to study the DNA distribution of mouse granulocyte-macrophage colony forming cells (GM-CFc) grown in a methyl cellulose culture system. This method involved the collection of cell clusters and c
Separation of subpopulations of in vitro colony forming cells from mouse marrow by equilibrium density centrifugation
β Scribed by Neil Williams; Gerrit J. Den Van Engh
- Publisher
- John Wiley and Sons
- Year
- 1975
- Tongue
- English
- Weight
- 610 KB
- Volume
- 86
- Category
- Article
- ISSN
- 0021-9541
No coin nor oath required. For personal study only.
β¦ Synopsis
Abstract
Equilibrium density centrifugation was used to characterise and separate subpopulations of mouse haemopoietic progenitor cells capable of producing colonies of granulocytes and macrophages in vitro. The material used to induce colony formation (CSF) was prepared from an extract of pregnant mouse uteri. This CSF preparation was found to be free of factors modifying the response. Under these culture conditions, in vitro colony forming cells (CFUβc) were found to be relatively homogeneous in their buoyant density. This homogeneity was independent of CSF concentration. A heterogeneous density profile of CFUβc was obtained when various cell fractions were cultured in the presence of CSF and rat blood lysate. The majority of the additional cells which responded to erythrocyte lysate were dense (modal density 1.080 g/cm^3^) compared to CFUβc which respond to CSF alone (modal density 1.074 g/cm^3^). It is concluded that in vitro colonies induced by CSF and in vitro colonies grown in the presence of CSF and erythrocyte lysate reflect two different populations of CFUβc.
π SIMILAR VOLUMES
Hemopoietic colony formation in agar occurred spontaneously in mass cultures of marrow cells obtained from a number of species (guinea pig, rat, lamb, rabbit, pig, calf, human and Rhesus monkey). This contrastcd with the observation that colony formation by mouse bone marrow exhibited an absolute re
## Abstract The technique of buoyant density separation in gradients of Bovine Serum Albumin has been used to separate hemopoietic cell populations in mouse bone marrow that form __in vivo__ spleen colonies and __in vitro__ colonies of granulocytes and macrophages in an agar culture system. The den
## Abstract C~57~BL bone marrow cells were separated on the basis of their sedimentation velocity at unit gravity and cell fractions cultured in agar using three types of colony stimulating factor (CSF). Colonyβforming cells separated as a single peak (s = 4.4 mm/hr) in cultures stimulated by mouse
Using a modification of the agar gel method for bone marrow culture, serum from various strains of mice has been tested for colony stimulating activity. Ninety percent of sera from AKR mice with spontaneous or transplanted lymphoid leukemia and 40-50% of sera from normal or preleukemic AKR mice sti
## Abstract In vitro macrophage colonyβforming cells (MβCFC) have been detected in bone marrow (BM) (317/10^5^ cells), spleen (SPL) (81/10^5^), and peripheral blood leukocytes (PBL) (242/10^5^) of the mouse. These MβCFCs were similar to those previously detected in thymus (T) (30/10^6^) and lymph n