# US 2013/0251684 A1 · Methods of isolating and culturing stem cells > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2013/0251684 A1 - Document ID: US20130251684A1 - Status: Published application - Kind: A1 - Application number: 13/782,968 - Filed: 2013-03-01 - Published: 2013-09-26 - Earliest claimed: 2012-03-02 - Assignees: Tissuetech, Inc. - Applicants: Tissuetech, Inc. - Inventors: Scheffer Tseng; Szu-Yu Chen; Suzhen Zhang - Web page: https://patentgrep.com/patent/US20130251684A1 ## CPC - C12N 5/0607: {Non-embryonic pluripotent stem cells, e.g. MASC (induced pluripotent stem cells C12N5/0696)} - C12N 5/0665: {Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood} - C12N 5/0667: {Adipose-derived stem cells [ADSC]; Adipose stromal stem cells} - C12N 2500/34: Sugars - C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2) - C12N 2501/235: Leukemia inhibitory factor [LIF] - C12N 2501/40: Regulators of development - C12N 2501/998: Proteins not provided for elsewhere - C12N 2533/80: Hyaluronan - C12N 2533/90: Substrates of biological origin, e.g. extracellular matrix, decellularised tissue - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Provided herein are methods of isolating and expanding a plurality of multipotent stem cells. Also described are methods of expanding stem cells on a substrate comprising an HC-HA complex. Also described are isolated and expanded stem cells produced by the methods and uses thereof, including stem cell therapy, as niche cells for supporting other types of stem cells, or as bioreactors for the production of HC-HA complexes. Also described are uses of HC-HA complexes as a carrier for stem cells. ## Claims ### Claim 1 A method for isolating an E-cadherin positive stem cell, comprising contacting a mixed cell population comprising one or more stem cells with an agent that binds to E-cadherin, thereby isolating an E-cadherin positive stem cell. ### Claim 2 (depends on 1) The method of claim 1, wherein the mixed cell population is substantially free of epithelial cells. ### Claim 3 (depends on 1) The method of claim 1, further comprising removing one or more epithelial cells from the mixed cell population. ### Claim 4 (depends on 1) The method of claim 1, further comprising removing one or more epithelial cells from the mixed cell population prior to contacting the mixed cell population with the agent. ### Claim 5 (depends on 1) The method of claim 1, wherein the agent is an antibody. ### Claim 6 (depends on 1) The method of claim 1, further comprising isolating the E-cadherin positive stem cell by fluorescence activated cell sorting or magnetic activated cell sorting. ### Claim 7 (depends on 1) The method of claim 1, wherein the mixed cell population comprises an embryonic stem cell, an adult stem cell, a fetal stem cell, or an induced pluripotent stem cell. ### Claim 8 (depends on 1) The method of claim 1, wherein the mixed cell population comprises a limbal stromal niche cell, an umbilical cord stem cell, an amniotic membrane stem cell or an adipose stem cell. ### Claim 9 (depends on 1) The method of claim 1, further comprising deriving the mixed cell population from an umbilical cord. ### Claim 10 (depends on 9) The method of claim 9, wherein the umbilical cord is a human, non-human primate, cow or pig umbilical cord. ### Claim 11 (depends on 9) The method of claim 9, further comprising: (a) mechanically or enzymatically removing the amniotic membrane epithelial cells from an umbilical cord, thereby producing remaining umbilical cord tissue; and (b) contacting the remaining umbilical cord tissue with collagenase for a period of time sufficient to separate one or more stem cells from other bound cells and components of the stromal matrix of the remaining umbilical cord tissue. ### Claim 12 (depends on 11) The method of claim 11, further comprising removing an umbilical cord blood vessel from the remaining umbilical cord tissue prior to digestion of the remaining umbilical cord tissue. ### Claim 13 (depends on 1) The method of claim 1, further comprising deriving the mixed cell population from adipose tissue. ### Claim 14 (depends on 13) The method of claim 13, further comprising digesting the adipose tissue with collagenase, thereby producing collagenase-digested adipose tissue. ### Claim 15 (depends on 14) The method of claim 14, further comprising digesting the adipose tissue in modified ESC medium. ### Claim 16 (depends on 14) The method of claim 14, further comprising fractionating the collagenase-digested adipose tissue by centrifugation, thereby producing a floating cell fraction (FC) and a sedimented stromal vascular fraction (SVF). ### Claim 17 (depends on 16) The method of claim 16, further comprising selecting the FC as the mixed cell population. ### Claim 18 (depends on 16) The method of claim 16, further comprising selecting the sedimented SVF as the mixed cell population. ### Claim 19 (depends on 16) The method of claim 16, further comprising filtering the sedimented SVF on a mesh filter, thereby producing a filtered SVF and a remaining cell fraction (RC) remaining on the filter. ### Claim 20 (depends on 19) The method of claim 19, further comprising selecting the filtered SVF as the mixed cell population. ### Claim 21 (depends on 19) The method of claim 19, further comprising selecting the RC as the mixed cell population. ### Claim 22 (depends on 19) The method of claim 19, wherein the filter has a pore size of about 40 μm to about 250 μm. ### Claim 23 (depends on 1) The method of claim 1, further comprising deriving the mixed cell population from amniotic membrane. ### Claim 24 (depends on 23) The method of claim 23, further comprising: (a) contacting the amniotic membrane with collagenase, thereby producing collagenase-digested amniotic membrane; and (b) contacting the collagenase-digested amniotic membrane with dispase. ### Claim 25 (depends on 23) The method of claim 23, further comprising: (a) contacting the amniotic membrane with dispase, thereby producing dispase-digested amniotic membrane; and (b) contacting the dispase-digested amniotic membrane with collagenase. ### Claim 26 (depends on 1) A method for expanding a stem cell population comprising: expanding one or more isolated stem cells of claim 1 in a culture comprising a two-dimensional substrate, thereby forming a plurality of expanding stem cells. ### Claim 27 (depends on 26) The method of claim 26, wherein at least one of the expanding stem cells does not pass the Hayflick limit. ### Claim 28 (depends on 26) The method of claim 26, wherein the two-dimensional substrate comprises Matrigel, laminin, fibronectin, collagen or entactin. ### Claim 29 (depends on 26) The method of claim 26, wherein the two-dimensional substrate comprises an HC-HA complex. ### Claim 30 (depends on 29) The method of claim 29, wherein the HC-HA complex is immobilized. ### Claim 31 (depends on 29) The method of claim 29, wherein the HC-HA complex is a native HC-HA complex or is a reconstituted HC-HA complex. ### Claim 32 (depends on 29) The method of claim 29, wherein the native HC-HA complex is an amniotic membrane HC-HA complex. ### Claim 33 (depends on 29) The method of claim 29, wherein the HC-HA complex comprises TSG-6. ### Claim 34 (depends on 29) The method of claim 29, wherein the HC-HA complex comprises PTX3. ### Claim 35 (depends on 29) The method of claim 29, wherein the HC-HA complex comprises a small leucine-rich proteoglycan. ### Claim 36 (depends on 35) The method of claim 35, wherein the small leucine-rich proteoglycan is selected from among decorin, biglycan, fibromodulin, lumican, PRELP (proline arginine rich end leucine-rich protein), keratocan, osteoadherin, epipycan, and osteoglycin. ### Claim 37 (depends on 29) The method of claim 29, wherein the HC-HA complex comprises TSG-6, PTX3, and a SLRP. ### Claim 38 (depends on 26) The method of claim 26, wherein the culture comprises supplemental hormonal epithelial media or embryonic stem cell media. ### Claim 39 (depends on 26) The method of claim 26, wherein the culture comprises bFGF or LIF. ### Claim 40 (depends on 26) The method of claim 26, wherein the culture comprises an inhibitor of Rho-associated kinase (ROCK). ### Claim 41 (depends on 26) The method of claim 26, further comprising (a) isolating at least one expanding stem cell from the plurality of expanding stem cells, thereby producing an isolated expanded stem cell; and (b) culturing the isolated expanded stem cell in a second culture comprising a three-dimensional substrate. ### Claim 42 (depends on 41) The method of claim 41, wherein the three-dimensional substrate comprises Matrigel, laminin, fibronectin, collagen or entactin. ### Claim 43 (depends on 41) The method of claim 41, wherein the second culture comprises supplemental hormonal epithelial media or embryonic stem cell media. ### Claim 44 A method for expanding a stem cell population comprising: expanding one or more stem cells in a culture comprising a two-dimensional substrate comprising an HC-HA complex, thereby forming a plurality of expanding stem cells. ### Claim 45 (depends on 44) The method of claim 44, wherein at least one of the expanding stem cells does not pass the Hayflick limit. ### Claim 46 (depends on 44) The method of claim 44, wherein the HC-HA complex is immobilized. ### Claim 47 (depends on 44) The method of claim 44, wherein the HC-HA complex is a native HC-HA complex or is a reconstituted HC-HA complex. ### Claim 48 (depends on 44) The method of claim 44, wherein the native HC-HA complex is an amniotic membrane HC-HA complex. ### Claim 49 (depends on 48) The method of claim 48, further comprising purifying the native HC-HA complex by performing ultracentrifugation on an amniotic membrane extract or an umbilical cord extract. ### Claim 50 (depends on 49) The method of claim 49, further comprising performing two, three or four rounds of ultracentifugation. ### Claim 51 (depends on 49) The method of claim 49, further comprising performing four rounds of ultracentifugation. ### Claim 52 (depends on 44) The method of claim 44, wherein the HC-HA complex comprises TSG-6. ### Claim 53 (depends on 44) The method of claim 44, wherein the HC-HA complex comprises PTX3. ### Claim 54 (depends on 44) The method of claim 44, wherein the HC-HA complex comprises a small leucine-rich proteoglycan. ### Claim 55 (depends on 54) The method of claim 54, wherein the small leucine-rich proteoglycan is selected from among decorin, biglycan, fibromodulin, lumican, PRELP (proline arginine rich end leucine-rich protein), keratocan, osteoadherin, epipycan, and osteoglycin. ### Claim 56 (depends on 44) The method of claim 44, wherein the HC-HA complex comprises TSG-6, PTX3, and a SLRP. ### Claim 57 (depends on 44) The method of claim 44, wherein the stem cell is an embryonic stem cell, an adult stem cell, a fetal stem cell, or an induced progenitor cell. ### Claim 58 (depends on 44) The method of claim 44, wherein the stem cell is a limbal stromal niche cell, an umbilical cord stem cell, an amniotic membrane stem cell or an adipose stem cell. ### Claim 59 (depends on 44) The method of claim 44, wherein the stem cell is a mesenchymal stem cell. ### Claim 60 (depends on 44) The method of claim 44, wherein the stem cell is a stem cell derived from an adult differentiated cell. ### Claim 61 (depends on 44) The method of claim 44, wherein the stem cell is a stem cell derived from a fibroblast. ### Claim 62 (depends on 61) The method of claim 61, wherein the stem cell is derived from a conjunctivochalasis fibroblast. ### Claim 63 (depends on 44) The method of claim 44, further comprising deriving the mixed cell population from an umbilical cord. ### Claim 64 (depends on 63) The method of claim 63, wherein the umbilical cord is a human, non-human primate, cow or pig umbilical cord. ### Claim 65 (depends on 63) The method of claim 63, further comprising: (a) mechanically or enzymatically removing the amniotic membrane epithelial cells from an umbilical cord, thereby producing remaining umbilical cord tissue; and (b) contacting the remaining umbilical cord tissue with collagenase for a period of time sufficient to separate one or more stem cells from other bound cells and components of the stromal matrix of the remaining umbilical cord tissue. ### Claim 66 (depends on 65) The method of claim 65, further comprising removing an umbilical cord blood vessel from the remaining umbilical cord tissue prior to digestion of the remaining umbilical cord tissue. ### Claim 67 (depends on 44) The method of claim 44, further comprising deriving the mixed cell population from adipose tissue. ### Claim 68 (depends on 67) The method of claim 67, further comprising digesting the adipose tissue with collagenase, thereby producing collagenase-digested adipose tissue. ### Claim 69 (depends on 68) The method of claim 68, further comprising digesting the adipose tissue in modified ESC medium. ### Claim 70 (depends on 68) The method of claim 68, further comprising fractionating the collagenase-digested adipose tissue by centrifugation, thereby producing a floating cell fraction (FC) and a sedimented stromal vascular fraction (SVF). ### Claim 71 (depends on 70) The method of claim 70, further comprising selecting the FC as the mixed cell population. ### Claim 72 (depends on 70) The method of claim 70, further comprising selecting the sedimented SVF as the mixed cell population. ### Claim 73 (depends on 70) The method of claim 70, further comprising filtering the sedimented SVF on a mesh filter, thereby producing a filtered SVF and a remaining cell fraction (RC) remaining on the filter. ### Claim 74 (depends on 73) The method of claim 73, further comprising selecting the filtered SVF as the mixed cell population. ### Claim 75 (depends on 73) The method of claim 73, further comprising selecting the RC as the mixed cell population. ### Claim 76 (depends on 73) The method of claim 73, wherein the filter has a pore size of about 40 μm to about 250 μm. ### Claim 77 (depends on 44) The method of claim 44, further comprising deriving the mixed cell population from amniotic membrane. ### Claim 78 (depends on 77) The method of claim 77, further comprising: (a) contacting the amniotic membrane with collagenase, thereby producing collagenase-digested amniotic membrane; and (b) contacting the collagenase-digested amniotic membrane with dispase. ### Claim 79 (depends on 77) The method of claim 77, further comprising: (a) contacting the amniotic membrane with dispase, thereby producing dispase-digested amniotic membrane; and (b) contacting the dispase-digested amniotic membrane with collagenase. ### Claim 80 (depends on 44) The method of claim 44, wherein the two-dimensional substrate comprises Matrigel, laminin, fibronectin, collagen or entactin. ### Claim 81 (depends on 44) The method of claim 44, wherein the culture comprises supplemental hormonal epithelial media or embryonic stem cell media. ### Claim 82 (depends on 44) The method of claim 44, wherein the culture comprises bFGF or LIF. ### Claim 83 (depends on 44) The method of claim 44, wherein the culture comprises an inhibitor of Rho-associated kinase (ROCK). ### Claim 84 (depends on 44) The method of claim 44, further comprising (a) isolating at least one expanding stem cell from the plurality of expanding stem cells, thereby producing an isolated expanded stem cell; and (b) culturing the isolated expanded stem cell in a second culture comprising a three-dimensional substrate. ### Claim 85 (depends on 84) The method of claim 84, wherein the three-dimensional substrate comprises Matrigel, laminin, fibronectin, collagen or entactin. ### Claim 86 (depends on 84) The method of claim 84, wherein the second culture comprises supplemental hormonal epithelial media or embryonic stem cell media. ### Claim 87 A method for expanding a stem cell comprising: (a) separating a plurality of cells of a tissue sample from components of an extracellular matrix in the tissue sample, to form a mixed cell population; (b) culturing the mixed cell population in a first culture comprising supplemented hormonal epithelial medium (SHEM) on a plastic tissue culture dish, thereby producing a population of non-adherent cells; (c) isolating the population of non-adherent cells; and (d) expanding at least one cell of the population of non-adherent cells in a second culture comprising a two-dimensional substrate, thereby forming a plurality of expanding stem cells. ### Claim 88 (depends on 87) The method of claim 87, wherein at least one of the expanding stem cells does not pass the Hayflick limit. ### Claim 89 (depends on 87) The method of claim 87, wherein the tissue is an amniotic membrane, an umbilical cord, a limbal tissue or an adipose tissue. ### Claim 90 (depends on 87) The method of claim 87, wherein the stem cell is a limbal stromal niche cell, an umbilical cord stem cell, an amniotic membrane stem cell or an adipose stem cell. ### Claim 91 (depends on 87) The method of claim 87, wherein the stem cell is a mesenchymal stem cell. ### Claim 92 (depends on 87) The method of claim 87, wherein the two-dimensional substrate comprises Matrigel, laminin, fibronectin, collagen or entactin. ### Claim 93 (depends on 87) The method of claim 87, wherein the two-dimensional substrate comprises an HC-HA complex. ### Claim 94 (depends on 93) The method of claim 93, wherein the HC-HA complex is a native HC-HA complex or is a reconstituted HC-HA complex. ### Claim 95 (depends on 94) The method of claim 94, wherein the native HC-HA complex is an amniotic membrane HC-HA complex. ### Claim 96 (depends on 95) The method of claim 95, further comprising purifying the native HC-HA complex by performing ultracentrifugation on an amniotic membrane extract or an umbilical cord extract. ### Claim 97 (depends on 96) The method of claim 96, further comprising performing two, three or four rounds of ultracentifugation. ### Claim 98 (depends on 96) The method of claim 96, further comprising performing four rounds of ultracentifugation. ### Claim 99 (depends on 93) The method of claim 93, wherein the HC-HA complex comprises TSG-6. ### Claim 100 (depends on 93) The method of claim 93, wherein the HC-HA complex comprises PTX3. ### Claim 101 (depends on 93) The method of claim 93, wherein the HC-HA complex comprises a small leucine-rich proteoglycan. ### Claim 102 (depends on 101) The method of claim 101, wherein the small leucine-rich proteoglycan is selected from among decorin, biglycan, fibromodulin, lumican, PRELP (proline arginine rich end leucine-rich protein), keratocan, osteoadherin, epipycan, and osteoglycin. ### Claim 103 (depends on 93) The method of claim 93, wherein the HC-HA complex comprises TSG-6, PTX3, and a SLRP. ### Claim 104 (depends on 87) The method of claim 87, wherein the second culture comprises supplemental hormonal epithelial media or embryonic stem cell media. ### Claim 105 (depends on 87) The method of claim 87, wherein the second culture comprises bFGF or LIF. ### Claim 106 (depends on 87) The method of claim 87, wherein the second culture comprises an inhibitor of Rho-associated kinase (ROCK). ### Claim 107 (depends on 87) The method of claim 87, wherein the tissue is an amniotic membrane. ### Claim 108 (depends on 107) The method of claim 107, further comprising: (i) contacting the amniotic membrane with collagenase, thereby producing collagenase-digested amniotic membrane; and (ii) contacting the collagenase-digested amniotic membrane with dispase. ### Claim 109 (depends on 107) The method of claim 107, further comprising: (i) contacting the amniotic membrane with dispase, thereby producing dispase-digested amniotic membrane; and (ii) contacting the dispase-digested amniotic membrane with collagenase. ### Claim 110 (depends on 87) The method of claim 87, further comprising: (a) isolating at least one expanding stem cell from the plurality of expanding stem cells, thereby producing an isolated expanding stem cell; and (b) culturing the isolated expanding stem cell in a second culture comprising a three-dimensional substrate. ### Claim 111 (depends on 109) The method of claim 109, wherein the three-dimensional substrate comprises Matrigel, laminin, fibronectin, collagen or entactin. ### Claim 112 (depends on 1, 26, 44) The method of any of claim 1, 26, 44, or 87, wherein the isolated or expanding stem cell is an amniotic membrane stem cell, and the method further comprises isolating an HC-HA complex from the stem cell. ### Claim 113 A method for inducing or maintaining pluripotency in a cell, comprising culturing the cell with an HC-HA complex, thereby inducing or maintaining pluripotency in the cell. ### Claim 114 (depends on 113) The method of claim 113, wherein the cell heterogeneously expresses a protein selected from among Sox2, myc, Oct4 and KLF4. ### Claim 115 (depends on 113) The method of claim 113, wherein the cell heterogeneously expresses one, two, or three proteins selected from among Sox2, myc, Oct4 and KLF4. ### Claim 116 (depends on 113) The method of claim 113, wherein the HC-HA complex is immobilized. ### Claim 117 (depends on 113) The method of claim 113, wherein the cell is an adult differentiated cell. ### Claim 118 (depends on 113) The method of claim 113, wherein the cell is a fibroblast. ### Claim 119 (depends on 118) The method of claim 118, wherein the cell is a conjunctivochalasis fibroblast. ### Claim 120 (depends on 113) The method of claim 113, wherein the cell is an embryonic stem cell, an adult stem cell, a fetal stem cell, or an induced pluripotent stem cell. ### Claim 121 (depends on 113) The method of claim 113, wherein the cell is a limbal epithelial progenitor cell, a limbal stromal niche cell, an umbilical cord stem cell, an amniotic membrane stem cell or an adipose stem cell. ### Claim 122 (depends on 113) The method of claim 113, wherein the HC-HA complex is a native HC-HA complex or is a reconstituted HC-HA complex. ### Claim 123 (depends on 122) The method of claim 122, wherein the native HC-HA complex is an amniotic membrane HC-HA complex. ### Claim 124 (depends on 123) The method of claim 123, further comprising purifying the native HC-HA complex by performing ultracentrifugation on an amniotic membrane extract or an umbilical cord extract. ### Claim 125 (depends on 124) The method of claim 124, further comprising performing two, three or four rounds of ultracentifugation. ### Claim 126 (depends on 124) The method of claim 124, further comprising performing four rounds of ultracentifugation. ### Claim 127 (depends on 113) The method of claim 113, wherein the HC-HA complex comprises TSG-6. ### Claim 128 (depends on 113) The method of claim 113, wherein the HC-HA complex comprises PTX3. ### Claim 129 (depends on 113) The method of claim 113, wherein the HC-HA complex comprises a small leucine-rich proteoglycan. ### Claim 130 (depends on 129) The method of claim 129, wherein the small leucine-rich proteoglycan is selected from among decorin, biglycan, fibromodulin, lumican, PRELP (proline arginine rich end leucine-rich protein), keratocan, osteoadherin, epipycan, and osteoglycin. ### Claim 131 (depends on 113) The method of claim 113, wherein the HC-HA complex comprises TSG-6, PTX3, and a SLRP. ### Claim 132 A method for producing an isolated native HC-HA complex comprising: (a) culturing an amniotic membrane stem cell, thereby producing a cultured amniotic membrane stem cell; and (b) isolating an HC-HA complex from the cultured stem cell, thereby producing an isolated native HC-HA complex. ### Claim 133 (depends on 132) The method of claim 132, wherein the amniotic membrane is from a placenta or an umbilical cord. ### Claim 134 (depends on 132) The method of claim 132, wherein the stem cell is an amniotic membrane stem cell or an umbilical cord stem cell. ### Claim 135 (depends on 134) The method of claim 134, wherein the umbilical cord stem cell is from the umbilical cord stromal layer or Wharton's jelly layer. ### Claim 136 (depends on 1, 26, 44) A method of treatment of an individual in need of a stem cell therapy, comprising administering to the individual a plurality of isolated or expanded stem cells of any of claim 1, 26, 44, or 87. ### Claim 137 (depends on 136) The method of claim 136, wherein the individual has a disease or condition selected from among non-healing wounds, diabetes, arthritis, inflammatory bowel disease, Crohn's disease, myocardial infarction, stroke, traumatic brain injury, spinal cord injury, learning defects, Alzheimer's disease, Parkinson's disease, baldness, missing teeth, osteoarthritis, rheumatoid arthritis, muscular dystrophy, cancer, amyotrophic lateral sclerosis. ### Claim 138 (depends on 136) The method of claim 136, comprising transplanting the isolated or expanded stem cells into the bone marrow of the individual. ### Claim 139 (depends on 1, 26, 44) A method of expanding a stem cell comprising culturing the stem cell in the presence of one or more isolated or expanded stem cells of any of claim 1, 26, 44, or 87. ### Claim 140 (depends on 1, 26, 44) A composition comprising a plurality of expanded stem cells of any of claim 1, 26, 44, or 87 and a pharmaceutically acceptable excipient. ### Claim 141 A composition comprising: (a) a stem cell; (b) an HC-HA complex; and (c) a tissue culture plate. ### Claim 142 (depends on 141) The composition of claim 141, wherein the HC-HA complex is immobilized to the tissue culture plate. ### Claim 143 (depends on 1, 26, 44, 141) The composition of claim 141, wherein the stem cell is an isolated or expanded stem cell of any of claim 1, 26, 44, or 87. ### Claim 144 (depends on 141) The composition of claim 141, wherein the stem cell is an embryonic stem cell, an adult stem cell, a fetal stem cell, or an induced progenitor cell. ### Claim 145 (depends on 141) The composition of claim 141, wherein the stem cell is a limbal stromal niche cell, an umbilical cord stem cell, an amniotic membrane stem cell or an adipose stem cell. ### Claim 146 (depends on 141) The composition of claim 141, wherein the stem cell is a mesenchymal stem cell. ### Claim 147 (depends on 141) The composition of claim 141, wherein the stem cell is an induced pluripotent stem cell derived from an adult differentiated cell. ### Claim 148 (depends on 141) The composition of claim 141, wherein the stem cell is an induced pluripotent stem cell derived from a fibroblast. ### Claim 149 (depends on 141) The composition of claim 141, wherein the stem cell is an induced pluripotent stem cell derived from a conjunctivochalasis fibroblast. ### Claim 150 (depends on 141) The composition of claim 141, wherein the HC-HA complex is a native HC-HA complex or is a reconstituted HC-HA complex. ### Claim 151 (depends on 141) The composition of claim 141, wherein the HC-HA complex comprises TSG-6. ### Claim 152 (depends on 141) The composition of claim 141, wherein the HC-HA complex comprises PTX3. ### Claim 153 (depends on 141) The method of claim 141, wherein the HC-HA complex comprises a small leucine-rich proteoglycan. ### Claim 154 (depends on 153) The method of claim 153, wherein the small leucine-rich proteoglycan is selected from among decorin, biglycan, fibromodulin, lumican, PRELP (proline arginine rich end leucine-rich protein), keratocan, osteoadherin, epipycan, and osteoglycin. ### Claim 155 (depends on 141) The method of claim 141, wherein the HC-HA complex comprises TSG-6, PTX3, and a SLRP.