Methods for regulating the specific lineages of cells produced in a human hematopoietic cell culture
First Claim
1. A method for controlling cellular lineage development in an ex vivo human hematopoietic cellular system, comprising culturing human stem and/or progenitor cells found in the human hematopoietic system in liquid culture media containing hematopoietic growth factors and which is replaced at a rate which is either (i) substantially continuous and providing ex vivo human stem cell division and/or human progenitor cell expansion therein or (ii) equal to 50 to 100% daily replacement for a cell density of from 1×
- 104 to 1×
107 cells per ml of culture, while maintaining said culture under physiologically acceptable conditions and adjusting the concentration of said hematopoietic growth factors as follows;
IL-3 or GM-CSF at a concentration of 0.1 to 100 ng/ml/day, Epo at a concentration of 0.001 to 10 U/ml/day, steel factor at a concentration of 1-100 ng/ml/day, IL-1 at a concentration of 10-100 U/ml/3-5 days, IL-6, G-CSF, bFGF, IL-7, IL-8, IL-9, IL-10, IL-11, PDGF or EGF at a concentration of 1-100 ng/ml/day, and mixtures thereof, to select for enhanced production of a desired human hematopgietic cell type.
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Abstract
Methods, including culture media conditions, which provide for in vitro human stem cell division and/or the optimization of human hematopoietic progenitor cell cultures and/or increasing the metabolism or GM-CSF secretion or IL-6 secretion of human stromal cells and/or a method for assaying the effect of a substance or condition on a human hematopoietic cell population, and/or depleting the malignant cell or T-cell and B-cell content of a human hematopoietic cell population are disclosed. The methods rely on culturing human stem cells and/or human hematopoietic progenitor cells and/or human stromal cells in a liquid culture medium which is replaced, preferably perfused, either continuously or periodically, at a rate of 1 ml of medium per ml of culture per about 24 to about 48 hour period, and removing metabolic products and replenishing depleted nutrients while maintaining the culture under physiologically acceptable conditions. Optionally, growth factors are added to the culture medium. The disclosed culture conditions afford improved methods for bone marrow transplantation.
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Citations
57 Claims
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1. A method for controlling cellular lineage development in an ex vivo human hematopoietic cellular system, comprising culturing human stem and/or progenitor cells found in the human hematopoietic system in liquid culture media containing hematopoietic growth factors and which is replaced at a rate which is either (i) substantially continuous and providing ex vivo human stem cell division and/or human progenitor cell expansion therein or (ii) equal to 50 to 100% daily replacement for a cell density of from 1×
- 104 to 1×
107 cells per ml of culture, while maintaining said culture under physiologically acceptable conditions and adjusting the concentration of said hematopoietic growth factors as follows;
IL-3 or GM-CSF at a concentration of 0.1 to 100 ng/ml/day, Epo at a concentration of 0.001 to 10 U/ml/day, steel factor at a concentration of 1-100 ng/ml/day, IL-1 at a concentration of 10-100 U/ml/3-5 days, IL-6, G-CSF, bFGF, IL-7, IL-8, IL-9, IL-10, IL-11, PDGF or EGF at a concentration of 1-100 ng/ml/day, and mixtures thereof, to select for enhanced production of a desired human hematopgietic cell type. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52)
- 104 to 1×
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53. In a method for controlling the lineage development of a cultured human cellular composition using hematopoietic growth factors, the improvement comprising culturing a human hematopoietic cell composition comprising human stem and/or progenitor cells found in the human hematopoietic system in a liquid culture medium which is replaced at a rate which is either (i) substantially continuous sufficient to obtain ex vivo human stem cell division and/or human progenitor cell expansion therein or (ii) equal to 50 to 100% daily replacement for a cell density of from 1×
- 104 to 1×
107 cells per ml of culture. - View Dependent Claims (54, 55, 56, 57)
- 104 to 1×
Specification
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- Resources
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Current AssigneeBoard of Regents of the University of Michigan (University of Michigan)
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Original AssigneeBoard of Regents of the University of Michigan (University of Michigan)
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InventorsPalsson, Bernhard O., Clarke, Michael F., Emerson, Stephen G., Armstrong, R. Douglas
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Primary Examiner(s)Ziska, Suzanne E.
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Application NumberUS08/334,011Time in Patent Office944 DaysField of Search435/240.23, 435/240.1, 435/240.2, 435/70.1US Class Current435/372CPC Class CodesA61K 48/00 Medicinal preparations cont...C07K 14/535 Granulocyte CSF; Granulocyt...C07K 14/5403 IL-3C07K 14/5412 IL-6C07K 14/705 Receptors; Cell surface ant...C12N 15/85 for animal cellsC12N 15/86 Viral vectorsC12N 2500/34 SugarsC12N 2501/125 Stem cell factor [SCF], c-k...C12N 2501/14 Erythropoietin [EPO]C12N 2501/22 Colony stimulating factors ...C12N 2501/23 Interleukins [IL]C12N 2501/39 Steroid hormonesC12N 2502/1394 Bone marrow stromal cells; ...C12N 2502/99 genetically modified cellsC12N 2740/10043 viral genome or elements th...C12N 5/0641 ErythrocytesC12N 5/0642 Granulocytes, e.g. basopils...C12N 5/0647 Haematopoietic stem cells; ...
