Human liver progenitors
First Claim
49. A method of preparing a composition comprising a mixture of cells derived from human liver tissue, which mixture comprises an enriched population of human liver progenitors, the method comprising:
- (a) providing a cell suspension of human liver tissue comprising a mixture of cells of varying sizes, including immature cells and mature cells; and
(b) debulking the suspension based on cell size, buoyant density, or a combination thereof to remove mature cells, while retaining immature cells, to provide a mixture of cells comprised of an enriched population of human liver progenitors.
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Abstract
Methods of isolating and cryopreserving progenitors from human liver are disclosed which include processing human liver tissue to provide a substantially single cell suspension comprising progenitors and non-progenitors of one or more cell lineages found in human liver; subjecting the suspension to a debulking step, which reduces substantially the number of non-progenitors in the suspension, and which provides a debulked suspension enriched in progenitors exhibiting one or more markers associated with at least one of the one or more cell lineages; and selecting from said debulked suspension those cells, which themselves, their progeny, or more mature forms thereof express one or more markers associated with at least one of the one or more cell lineages. Among these markers are CD14, CD34, CD38, CD45, and ICAM. Hepatic progenitors are characterized as being 6-15μ in diameter, diploid, glycophorin A−, CD45−, AFP+++, ALB+, ICAM+, and with subpopulations varying in expression of CD14+. CD34++, CD38++, CD117+. These progenitor subpopulations have characteristics expected for cells that are particularly useful in liver cell and gene therapies and for establishing bioartificial organs.
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Citations
68 Claims
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49. A method of preparing a composition comprising a mixture of cells derived from human liver tissue, which mixture comprises an enriched population of human liver progenitors, the method comprising:
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(a) providing a cell suspension of human liver tissue comprising a mixture of cells of varying sizes, including immature cells and mature cells; and
(b) debulking the suspension based on cell size, buoyant density, or a combination thereof to remove mature cells, while retaining immature cells, to provide a mixture of cells comprised of an enriched population of human liver progenitors. - View Dependent Claims (50, 51, 52, 53, 54, 55, 56, 65, 66, 67, 68)
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57. A method of preparing a composition comprising an enriched population of human hepatic progenitors comprising:
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(a) providing a cell suspension of human liver tissue, (b) debulking the suspension based on cell size, buoyant density, or a combination thereof to remove mature cells, and (c) subjecting the debulked suspension to a positive or negative immunoselection, such that a mixture of cells is provided, which mixture of cells is comprised of an enriched population of human hepatic progenitors, which human hepatic progenitors themselves, their progeny, or more mature forms thereof exhibit one or more markers indicative of expression of alpha-fetoprotein, albumin, or both. - View Dependent Claims (58, 59, 60, 61, 62, 63)
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64. A cell culture comprising an enriched population of human hepatic pluripotent progenitors, their progeny, or more mature forms thereof, which exhibit one or more markers indicative of expression of full-length alpha-fetoprotein, full-length albumin, or both, an extracellular matrix component, and a culture medium.
Specification