Differentiation of subpopulations of human and murine hemopoietic stem cells by hypotonic lysis.

Niskanen, Eero; Cline, Martin J.

doi:10.1172/jci109670

Cited by 20 publications

(7 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, in these experiments bone marrow cells were first separated in an albumin gradient and only the cells with a density lower than 1.070 g/ml were used for velocity sedimentation. Therefore, a majority of those CFU-D with a density above 1.068 g/ml that we have found in Percoll gradients were not included in the velocity sedimentation experiments, which may have contributed to the seemingly homogeneous size distribution of Our findings indicate non-identity between CFU-D and CFU-GM, in agreement with previous studies (12,13,20), and emphasize the fact that CFU-D is a heterogeneous population (23,24). Some of the colonies in diffusion chambers, and especially those derived from cells of medium to high density, arise from cells of intermediate maturity such as promyelocytes and marrow monocytes, and not necessarily from primitive CFU-D's with stem cell properties.…”

Section: Discussionsupporting

confidence: 89%

See 1 more Smart Citation

Density distribution of chronic myeloid leukaemia and normal colony‐forming cells in diffusion chambers (CFU‐D) and agar (CFU‐GM)

Nilsson

Olofsson

1985

Scandinavian Journal of Haematology

View full text Add to dashboard Cite

The density distribution in Percoll gradients of clonogenic cells forming colonies in diffusion chambers (CFU‐D) or in agar culture (CFU‐GM) was studied in chronic myeloid leukaemia (CML). The density distribution of CFU‐GM in CML was homogeneous with peaks within 1.058–1.061 g/ml, which is slightly lower than normal. CFU‐D, on the other hand, showed heterogeneous distributions both in CML and in normal controls. Two separable populations of CFU‐D were recognized, one with the same or lower density than CFU‐GM that formed almost exclusively neutrophilic colonies in diffusion chambers, and a second population concentrating in the density range 1.068–1.075 g/ml which primarily formed macrophage colonies. The second population contained colony‐forming cells derived from both Fc‐receptor positive and Fc‐receptor negative precursor cells, suggesting that at least some colonies in diffusion chambers arise from Fc‐receptor positive granulopoietic cells of intermediate maturity and/or monocytes. The concentration of CFU‐D in peripheral blood was increased 40‐ to 100‐fold in 2 patients currently off treatment who had increased WBC counts.

show abstract

Section: Discussionsupporting

confidence: 89%

“…There is evidence that CFU-D is an intermediate stage between CFU-S and CFU-GM (12,13,20,21) and a possible parent-progeny relationship between CFU-D and CFU-GM has been demonstrated (22). In velocity sedimentation at 1 g CFU-D is a smaller cell than CFU-GM (13) with a relatively homogeneous size distribution.…”

Section: Discussionmentioning

confidence: 99%

Density distribution of chronic myeloid leukaemia and normal colony‐forming cells in diffusion chambers (CFU‐D) and agar (CFU‐GM)

Nilsson

Olofsson

1985

Scandinavian Journal of Haematology

View full text Add to dashboard Cite

show abstract

“…In preliminary experiments designed to e.stablish optimal conditions for Countcrllow Centrifugation manipulation of foetal calf .serum concentration, loading How rate or cell load (data not shown) did not prevent clumping of the eells and formation of a pellet in the elutriator chamber, and. although Meyskens et al (1979) did establish that hypotonic lysis of bone marrow cells resulted in improved separation due to removal of erythrocytes, Niskanen and Cline (1980) have shown that this can also result in loss of agar colony-forming cells. Since we were interested in evaluating the two methods for recovery and .separation of functional stem cells, we did not consider this procedure advisable.…”

Section: Discussionmentioning

confidence: 99%

A Comparison of Counterflow Centrifugation and Unit Gravity Sedimentation in the Separation of Mouse Bone Marrow Progenitor Cells

Attwood¹,

Bertoncello²,

Bradley³

et al. 1982

Aust J Exp Biol Med

View full text Add to dashboard Cite

Summary Both Unit Gravity Sedimentation and Counterflow Centrifugation separate cells primarily on the basis of size, However, when performed concurrently on the same normal mouse bone marrow cell suspensions, Unit Gravity Sedimentation was found to be far superior in terms of the recovery of total nucleated tell loud and the recovery and enrichment of two classes of functional progenitors: (1)those responsive to the combined stimulus of pregnant mouse uterus extract (PMUE) plus human spleen conditioned medium (HUSPCM), and (2), those responsive to PMUE alone. Neither technique was able to resolve PMUE from PMUE + HUSPCM progenitors. The major limitation of Counterflow Centrifugatton for the separation of sub‐populations of marrow cells is related to the complexity and range of cell sizes in marrow cell suspensions which lead to significant pellet formation within the elutriator chamber. Furthermore, cells undergoing Counterflow Centrifugation are subjected to forces not encountered in Unit Gravity Sedimentation. Therefore, the basis of the separations is not identical and this is reflected in cell volume distributions of comparable fractions separated by each method.

show abstract

“…Initially, based on morphological evaluation, they were considered identical (or closely related) to cells giving rise to CFU-gm in vitro [38]. However, in other studies these two precursors have been distinguished by velocity sedimentation [39, 401, density [41], cell cycle status [26,[42][43][44], resistance to osmotic lysis [45], sensitivity to thermal challenges [46,47], growth kinetics [23,48] and by the difference in response to growth factors [24,49,501. The relationship between CFU-d and other colony-forming progenitors is less clear because they often have similar characteristics.…”

Section: Introductionmentioning

confidence: 99%

Diffusion chamber colony‐forming unit (cfu‐d): A primitive stem cell

Niskanen¹,

Chatelain²,

Symann³

1989

The International Journal of Cell Cloning

View full text Add to dashboard Cite

Assay of hematopoietic precursor cells in diffusion chambers (DCs) implanted intraperitoneally in experimental animals provides a powerful tool for studying stem cell kinetics in vivo. In this system, the effect of cell migration (which complicates whole animal studies) is eliminated because the membranes utilized in the construction of the chambers are impermeable for cells, while permitting free passage of molecules present in the humoral phase of the host. As judged by light microscopy, conditions in the DC cultures primarily favor macrophage and granulocyte growth. However, the use of in vitro and in vivo subculture to further analyze chamber contents has demonstrated that the system supports proliferation of early hematopoietic progenitors. Additionally, cells capable of rescuing lethally irradiated mice proliferate in DC cultures. Development of the plasma clot DC technique has revealed that most of the growth occurs in colonies which are derived from single cells (CFU-d). Characterization of these cells indicates that they are at least as primitive as other colony-forming cells and, also based on subculture studies, can differentiate along several hematopoietic lineages. In addition to normal CFU-d, both embryonal and leukemic cells can give rise to granulocytes, macrophages, megakaryocytes and erythroid cells in the DC cultures. Evaluation of the effects of humoral factors on hematopoietic cell proliferation and differentiation in the system has led to the identification of both stimulators and inhibitors that may be different from the well-characterized cytokines. Thus, the system seems to be useful for detecting molecules controlling the most primitive stages of hematopoiesis. We believe that the DC culture technique holds enormous potential in the study of stem cell proliferation and differentiation in vivo.

show abstract

Differentiation of subpopulations of human and murine hemopoietic stem cells by hypotonic lysis.

Cited by 20 publications

References 17 publications

Density distribution of chronic myeloid leukaemia and normal colony‐forming cells in diffusion chambers (CFU‐D) and agar (CFU‐GM)

Density distribution of chronic myeloid leukaemia and normal colony‐forming cells in diffusion chambers (CFU‐D) and agar (CFU‐GM)

A Comparison of Counterflow Centrifugation and Unit Gravity Sedimentation in the Separation of Mouse Bone Marrow Progenitor Cells

Diffusion chamber colony‐forming unit (cfu‐d): A primitive stem cell

Contact Info

Product

Resources

About