Human hematopoietic stem and progenitor cell antigen and methods for its use
0 Assignments
0 Petitions
Accused Products
Abstract
A hematopoietic progenitor cell antigen and reagents, notably antibodies, that specifically bind to the antigen are provided. Expression of the antigen is highly tissue specific. It is only detected on a subset of hematopoietic progenitor cells derived from human bone marrow, fetal bone marrow and liver, cord blood and adult peripheral blood. The subset of cells recognized by AC133 is CD34bright and contains substantially all of the CFU-GM activity present in the CD34+ population. This highly specific distribution of AC133 makes it exceptionally useful as a reagent for isolating and characterizing human hematopoietic progenitor and stem cells. Cells selected for expression of AC133 antigen can be further purified by selection for other hematopoietic stem cell and progenitor cell markers.
15 Citations
51 Claims
- 1. An antibody that specifically binds to AC133 antigen.
-
6. A method for enrichment of hematopoietic stem or progenitor cells or both, said method comprising:
-
combining a mixed population of human cells comprising hematopoietic stem or progenitor cells or both with a reagent that specifically binds to the hematopoietic progenitor cell antigen recognized by the antibody produced by the hybridoma cell line ATCC ______; and
selecting for those cells that bind said reagent;
wherein said selected cells are enriched in hematopoietic stem or progenitor cell activity or both, depending on whether said mixed popluation of human cells contained hematopoietic stem or progenitor cells or both, respectively. - View Dependent Claims (7, 8, 9, 10, 11, 12, 35)
-
- 13. A substantially pure population of hematopoietic progenitor cells, wherein said cells are bound to a reagent that specifically binds to the hematopoietic progenitor cell antigen recognized by the antibody produced by the hybridoma cell line ATCC HB12346.
-
21. An isolated nucleic acid molecule, wherein said molecule comprises:
-
(1) a first sequence having an amino acid coding region for AC133 as set forth in FIG. 12 (SEQ ID NO;
1);
(2) a second sequence, wherein said second sequence is a subsequence of said first sequence and is at least 14 nucleotides in length;
(3) a third sequence in which at least one nucleotide of said first or second sequences is replaced by a different nucleotide;
or(4) a fourth sequence complementary to any of said first, second or third sequences;
with the proviso that (i) if said molecule is an RNA molecule, U replaces T in said sequence of said molecule, (ii) said third sequence is at least 90% identical to said first or second sequence, and (iii) said second sequence is not nucleotides 347-667, 1564-1696, or 2010-2386 of SEQ ID NO;
1. - View Dependent Claims (22, 23, 24, 25, 26, 27)
-
-
28. An isolated polypeptide, wherein said polypeptide comprises:
- (1) a first amino acid sequence of AC133 as set forth in SEQ ID NO;
2;
(2) a second amino acid sequence wherein said second sequence is a subsequence of said first sequences and is at least 6 amino acids in length;
or (3) a third sequence in which at least one amino acid of said first or second sequences is replaced by a different amino acid, with the proviso that said amino acid replacement is a replacement of one acidic residue for another, one basic residue for another, one non-polar residue for another, one uncharged polar residue for another, or one aromatic residue for another, with the proviso that said third sequence is at least 90% identical to said first or second sequence. - View Dependent Claims (29, 30, 31, 32, 33, 36, 37, 38, 39, 40, 48)
- (1) a first amino acid sequence of AC133 as set forth in SEQ ID NO;
-
34. An isolated polypeptide, wherein said polypeptide comprises the amino acid sequence from extracellular N-terminus, aa 20-107;
- first transmembrane region, aa 107-126;
first cytoplasmic loop, aa 127-157;
second transmembrane region, aa 158-179;
first extracellular loop, aa 180-435;
third transmembrane region, aa 436-454;
second cytoplasmic loop, aa 455-480;
fourth transmembrane region, aa 481-503;
second extracellular loop, aa 504-792;
fifth transmembrane, aa 793-816;
or cytoplasmic C-terminus, aa 817-865;
of SEQ ID NO;
2.
- first transmembrane region, aa 107-126;
-
41. In a method of isolating hematopoietic stem cells using a cell separation technique based on identification of stem cells by selective binding of a ligand to an antigenic marker on said stem cell, an improvement which comprises:
utilizing an AC133 antigen as said antigenic marker. - View Dependent Claims (42, 43, 44, 45, 46, 47)
- 49. A reagent that bind specifically to AC133 antigen with less than 5% crossreactivity with any antigen present on mature blood cells.
Specification