METHODS AND COMPOSITIONS FOR CAPTURE OF CELLS
Compositions and methods are provided for capturing cells. Cell binding peptides are provided that bind to one or more of stem cells, fibroblasts, or endothelial cells. In the methods, a sample containing cells is contacted with a cell binding peptide attached to a substrate, and the cells present in the sample are captured onto the substrate through binding to the cell binding peptide.
1 . A method for capturing cells, comprising contacting a sample comprising cells with a cell binding peptide attached to a substrate, wherein the cell binding peptide is selected from the group consisting of: SEQ ID NOs: 1-20, conservatively substituted variants of SEQ ID NOs: 1-16, and variants having at least 65% sequence identity to SEQ ID NOs: 1-16, and wherein the cells comprised in the sample are captured onto the substrate through binding to the cell binding peptide.
2 . The method of claim 1 , wherein the cell binding peptide binds to one or more of stem cells, fibroblasts, or endothelial cells and the sample comprising cells comprises one or more of stem cells, fibroblasts, or endothelial cells.
3 . The method of claim 1 , wherein the sample comprising cells comprises bone marrow, allogeneic stem cells, adipose tissue, stromal vascular fraction of adipose tissue, blood, blood products, platelets, platelet-rich plasma (PRP), umbilical cord blood, embryonic tissues, placenta, amniotic epithelial cells, tissue punch, omentum, or a homogeneous or heterogeneous population of cultured cells, or combinations or derivatives thereof.
4 . The method of claim 2 , further comprising a step of releasing the captured cells from the substrate.
5 . The method of claim 4 , wherein the captured cells are culture expanded on the substrate prior to release from the substrate.
6 . The method of claim 4 , wherein the step of releasing the captured stem cells is a physical means comprising shaking or centrifugation.
7 . The method of claim 4 , wherein the step of releasing the captured stem cells is a chemical means comprising a change in pH, a change in salt concentration, or a competitive inhibition binding with molecules that compete with the captured stem cells.
8 . The method of claim 1 , wherein the cell binding peptide is attached to the substrate through a spacer.
9 . The method of claim 8 , wherein the spacer is selected from the group consisting of amino acids, polymers, synthetic polymers, polyethers, poly(ethylene glycol) (“PEG”), An 11 unit polyethylene glycol (“PEG10”), and a 1 unit polyethylene glycol (“mini-PEG” or “MP”), and combinations thereof.
10 . The method of claim 1 , wherein the cell binding peptide comprises one or more modifications to the peptide N-terminus, peptide C-terminus, or within the peptide amino acid sequence, to allow for attachment of the cell binding peptide to the substrate or release of the cell binding peptide from the substrate.
11 . The method of claim 10 , wherein the modification is selected from the group consisting of amino acids, cysteine, lysine, hydroxyl group, thiol group, aldehyde group, acetyl group, polymers, synthetic polymers, polyethers, poly(ethylene glycol) (“PEG”), an 11 unit polyethylene glycol (“PEG10”), and a 1 unit polyethylene glycol (“mini-PEG” or “MP”), a group cleavable by addition of a reducing agent, an amino acid sequence cleavable by addition of an enzyme, and combinations thereof.
12 . The method of claim 1 , wherein the substrate comprises metal, glass, plastic, synthetic matrix, silica gel, polymer, polysaccharide, dextran, agarose, biopolymer, collagen, hydrogel, gel, keratin, silk, cellulose derivative, oxidized cellulose, oxidized regenerated cellulose, carboxymethylcellulose, hydroxypropylmethylcellulose, chitosan, chitin, hyaluronic acid or derivatives or combinations thereof.
13 . The method of claim 1 , wherein the substrate is in the form of beads, coated beads, gel, hydrogel, mesh, fibrous form, hollow fibers, or sheets.
14 . The method of claim 13 , wherein the cell capture is performed by an adsorption column, an adsorption membrane, or a density centrifugation.
15 . A device for chromatography comprising the a cell binding peptide attached to a substrate, wherein the cell binding peptide is selected from the group consisting of: SEQ ID NOs: 1-20, conservatively substituted variants of SEQ ID NOs: 1-16, and variants having at least 65% sequence identity to SEQ ID NOs: 1-16.
16 . A filter apparatus for capturing cells from a sample, the filter apparatus comprising a substrate to which a cell binding peptide is attached, wherein the cell binding peptide is selected from the group consisting of: SEQ ID NOs: 1-20, conservatively substituted variants of SEQ ID NOs: 1-16, and variants having at least 65% sequence identity to SEQ ID NOs: 1-16.
17 . A method for delivering cells to a subject, comprising:
a. contacting a sample comprising cells with a cell binding peptide attached to a substrate, wherein the cell binding peptide is selected from the group consisting of: SEQ ID NOs: 1-20, conservatively substituted variants of SEQ ID NOs: 1-16, and variants having at least 65% sequence identity to SEQ ID NOs: 1-16, wherein the cells comprised in the sample are captured onto the substrate through binding to the cell binding peptide;
b. releasing the cells from the cell binding peptide; and
c. delivering the released cells to the subject.
18 . A method for delivering cells to a subject, comprising contacting a sample comprising cells with a cell binding peptide attached to a substrate in the form of a bead, wherein the cell binding peptide is selected from the group consisting of: SEQ ID NOs: 1-20, conservatively substituted variants of SEQ ID NOs: 1-16, and variants having at least 65% sequence identity to SEQ ID NOs: 1-16, wherein the cells comprised in the sample are captured onto the bead substrate through binding to the cell binding peptide; and delivering the beads having the captured cells to the subject.
19 . A method for enhancing cell attachment to a synthetic matrix comprising, contacting a sample comprising cells with a collagen having a covalently attached cell binding peptide, wherein the cell binding peptide is selected from the group consisting of: SEQ ID NOs: 1-20, conservatively substituted variants of SEQ ID NOs: 1-16, and variants having at least 65% sequence identity to SEQ ID NOs: 1-16, wherein the cells comprised in the sample are captured onto the collagen through binding to the attached cell binding peptide; and contacting the collagen having the cells bound to the attached cell binding peptide with the synthetic matrix, wherein the attachment of the cells bound to the attached cell binding peptide to the synthetic matrix is enhanced relative to free cells.
20 . The method of claim 19 , wherein the synthetic matrix is selected from the group consisting of plastic, polypropylene, polystyrene, polytetrafluoroethylene (PTFE), expanded polytetrafluoroethylene (ePTFE), and polyether ether ketone (PEEK).
21 . The method of claim 19 , wherein the synthetic matrix is a cell culture plate.
22 . The method of claim 19 , wherein the cells are one or more of stem cells, fibroblasts, or endothelial cells.
23 . A method for promoting tissue integration of an implantable device, comprising:
a. coating the implantable device with a polymer having a covalently attached cell binding peptide;
b. contacting the coated implantable device with a sample comprising cells, wherein the cells comprised in the sample are captured onto the collagen substrate through binding to the attached cell binding peptide; and
c. delivering the coated implantable device having the bound cells to a subject for promoting tissue integration of the implantable device.
24 . The method of claim 23 , wherein the polymer is selected from the group consisting of a collagen, a keratin, a silk, a polysaccharide, an agarose, a cellulose derivative, an oxidized cellulose, an oxidized regenerated cellulose, a carboxymethylcellulose, a hydroxypropylmethylcellulose, a chitosan, a chitin, a hyaluronic acid, and derivatives and combinations thereof.
25 . The method of claim 23 , wherein the implantable device comprises one or a combination of plastic, polypropylene, polystyrene, polytetrafluoroethylene (PTFE), expanded polytetrafluoroethylene (ePTFE), polyether ether ketone (PEEK), or polyether ether ketone (PEEK).
26 . The method of claim 23 , wherein the polymer is collagen and the implantable device comprises polyether ether ketone (PEEK).