Method for creating perfusable microvessel systems

(54) METHOD FOR CREATING PERFUSABLE MICROVESSEL SYSTEMS

(75) Inventor: Thomas Neumann, Seattle, WA (U S)

(73) Assignee: Nortis, Inc., Gig Harbor, WA (U S)

( * ) Notice: Subject to any disclaimer, the term of tl1is

patent is extended or adjusted under 35 U.S.C. 154(b) by 916 days.

This patent is subject to a tenninal disclaimer.

(21) Appl.No.: 11/860,471 (22) Filed: Sep.24,2007

(65) Prior Publication Data US 2008/0261306 A1 Oct. 23, 2008

Related U.S. Application Data

(63) Continuation-in-part of application No. 11/388,920, filed on Mar. 24, 2006, now Pat. No. 7,622,298.

(51) Int. Cl. C12N 5/00 (2006.01) (52) U.S. Cl. ....................... .. 435/395; 435/325; 435/397 (58) Field of Classification Search ................ .. 435/395, 435/325, 397 See application file for complete search history. (56) References Cited U.S. PATENT DOCUMENTS 4,878,908 A 11/1989 Martin et al. 5,804,366 A 9/1998 Hu et al. 6,503,273 B1 1/2003 McAllister et al 6,592,623 B1 7/2003 Bowin et al. 6,642,019 B1 11/2003 Anderson et al. 6,893,812 B2 5/2005 Woltering 6,989,071 B2 1/2006 Kocur et al. 2002/0150879 A1 10/2002 Woltering et al. 2002/0177121 A1 11/2002 Woltering et al. 2003/0138945 A1 7/2003 McAllister et al 2003/0138950 A1 7/2003 McAllister et al. 2003/0171053 A1 9/2003 Sanders 2004/0044403 A1 3/2004 Bischoff et al. 2006/0216320 A1 9/2006 Kitazono et al. 2007/0110962 A1 5/2007 Tien et al. 2007/0224677 A1 9/2007 Neumann OTHER PUBLICATIONS

Nicolas L’Heureux et a1.,“A Completely biological tissue-engineered human blood vessel,” The FASEB Journal. vol. 12 (1), pp. 47-56, Jan. 1998.

Antonio Migliore et al. “Controlled in vitro growth of cell microtubes: towards the realisation of artificial microvessels,” Biomed Microdevices. vol. 10, pp. 81-88, Aug. 9, 2007.

Akhtar N, Dickerson EB, Auerbach R. 2002. The sponge/Matrigel angiogenesis assay. Angiogenesis 5:75-80.

Algire GH, Chalkley HW, Legallais FY, Park HD. 1945. Vascular reactions of normal and malignant tissues in vivo. I. Vascular reactions of mice to wounds and to normal and neoplastic transplants. J Natl Cancer Inst 6:73-85.

Andrade SP, Machado RD, Teixeira AS, Belo AV, Tarso AM, Beraldo WT. 1997. Sponge-induced angiogenesis in mice and the pharmaco

logical reactivity of the neovasculature quantitated by a fluorimetric method. Microvasc Res 54:253-261.

Arthur WT, Vernon RB, Sage EH, Reed MJ. 1998. Growth factors reverse the impaired sprouting of microvessels from aged mice. Microvasc Res 55:260-270.

Ausprunk DH, Knighton DR, Folkman J. 1974. Differentiation of vascular endothelium in the chick chorioallantois: a structural and autoradiographic study. Dev Biol 38:237-248.

Clark ER, Clark EL. 1939. Microscopic observations on the growth ofblood capillaries in the living mammal. Am J Anat 64:251-301. Davis GE, Camarillo CW. 1996.An alpha 2 beta 1 integrin-dependent pinocytic mechanism involving intracellular vacuole formation and coalescence regulates capillary lumen and tube formation in threedimensional collagen matrix. Exp Cell Res 224:39-51.

Elsdale T, Bard J. 1972. Collagen substrata for studies on cell behavior. J Cell Biol 54:626-637.

Feder J, Marasa J C, Olander JV. 1983. The formation of capillary-like tubes by calf aortic endothelial cells grown in vitro. J Cell Physiol 1 16: 1-6.

Fishman JA, Ryan GB, Karnovsky MJ. 1975. Endothelial regeneration in the rat carotid artery and the significance of endothelial denudation in the pathogenesis of myointimal thickening. Lab Invest 32:339-3 5 1.

Folkman J, Haudenschild C. 1980. Angiogenesis in vitro. Nature 288:551-556.

Folkman J , Haudenschild CC, Zetter BR. 1979. Long-term culture of capillary endothelial cells. Proc Natl Acad Sci U S A 76:5217-5221. Gimbrone MA, Jr. 1976. Culture of vascular endothelium. In: Prog Hemost Thromb. p. 1-28.

Gimbrone MA, Jr., Cotran RS, Folkman J . 1974a. Human vascular endothelial cells in culture. Growth and DNA synthesis. J Cell Biol 60:673-684.

Gimbrone MA, Jr., Cotran RS, Leapman SB, Folkman J . 1974b. Tumor growth and neovascularization: an experimental model using the rabbit cornea. J Natl Cancer Inst 52:413-427.

Greenblatt M, Shubi P. 1968. Tumor angiogenesis: transfilter diffusion studies in the hamster by the transparent chamber technique. J Natl Cancer Inst 41:111-124.

Hoying JB, Boswell CA, Williams SK. 1996. Angiogenic potential of microvessel fragments established in three-dimensional collagen gels. In Vitro Cell Dev Biol Anim 32:409-419.

Jaffe EA, Nachman RL, Becker CG, Minick CR. 1973. Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52:27452756.

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Primary Examiner — Ruth Davis (74) Attorney, Agent, or Firm — George A. Leone; Citadel Patent Law

(57) ABSTRACT

A method for creating networks of perfusable microves sels in vitro. A mandrel is drawn through a matrix to fonn a channel through the matrix. Cells are injected into the channel. The matrix is incubated to allow the cells to attach inside the channel. The channel is perfused to remove unattached cells to create a parent vessel, where the parent vessel includes a perfusable hollow channel lined with cells in the matrix. The parent vessel is induced to create sprouts into the surrounding matrix gel so as to form a microvessel network. The microvessel network is subjected to luminal perfusion through the parent vessel.

12 Claims, 10 Drawing Sheets

OTHER PUBLICATIONS

Jozaki K, Marucha PT, Despins AW, Kreutzer DL. 1990. An in vitro model of cell migration: evaluation of vascular endothelial cell migration. Anal Biochem 190139-47.

Koike T, Vernon RB, Gooden MD, Sadoun E, Reed MJ. 2003. Inhibited angiogenesis in aging: a role for TIMP-2. J Gerontol A Biol Sci Med Sci 58:B798-805.

Kubota Y, Kleinman HK, Martin GR, Lawley TJ. 1988. Role of laminin and basement membrane in the morphological differentiation of human endothelial cells into capillary-like structures. J Cell Biol 10711589-1598.

Kuzuya M, Kinsella JL. 1994. Induction of endothelial cell differentiation in vitro by fibroblast-derived soluble factors. Exp Cell Res 2151310-318.

Maciag T, Kadish J , Wilkins L, Stemerman MB, Weinstein R. 1982. Organizational behavior of human umbilical vein endothelial cells. J Cell Biol 94:511-520.

Madri JA. 1982. Endothelial cell-matrix interactions in hemostasis. Prog Hemost T11romb 6: 1-24.

Madri JA, Pratt BM. 1986. Endothelial cell-matrix interactions: in vitro models of angiogenesis. J Histochem Cytochem 34:85-91. Madri JA, Pratt BM, Tucker AM. 1988. Phenotypic modulation of endothelial cells by transforming growth factor-beta depends upon the composition and organization of the extracellular matrix. J Cell Biol 106:1375-1384.

Madri JA, Stenn KS. 1982. Aortic endothelial cell migration. I. Matrix requirements and composition. Am J Pathol 1061180-186. Manoussaki D, Lubkin SR, Vernon RB, Murray JD. 1996. A mechanical model for the formation of vascular networks in vitro. Acta Biotheor 44:271-282.

Marx M, Perlmutter RA, Madri JA. 1994. Modulation of plateletderived growth factor receptor expression in microvascular endothelial cells during in vitro angiogenesis. J Clin Invest 93:131139.

Merwin JR, Anderson J M, Kocher O, Van Itallie CM, Madri JA. 1990. Transforming growth factor beta 1 modulates extracellular matrix organization and cell-cell junctional complex formation during in vitro angiogenesis. J Cell Physiol 1421117-128.

Montesano R, Orci L. 1985. Tumor-promoting phorbol esters induce angiogenesis in vitro. Cell 42:469-477.

Montesano R, Orci L, Vassalli P. 1983. In vitro rapid organization of endothelial cells into capillary-like networks is promoted by collagen matrices. J Cell Biol 97: 1648-1652.

Montesano R, Pepper MS, Orci L. 1993. Paracrine induction of angiogenesis in vitro by Swiss 3T3 fibroblasts. J Cell Sci 105 ( Pt 4):1013-1024.

Mori M, SadahiraY, Kawasaki S, Hayashi T, Notohara K, Awai M. 1988. Capillary growth from reversed rat aortic segments cultured in collagen gel. Acta Pathol Jpn 38: 1503-1512.

Nehls V, Drenckhahn D. 1995. A novel, microcarrier-based in vitro assay for rapid and reliable quantification of three-dimensional cell migration and angiogenesis. Microvasc Res 50:311-322.

Nehls V, Herrmann R. 1996. The configuration of fibrin clots determines capillary morphogenesis and endothelial cell migration. Microvasc Res 51:347-364.

Neumann T, Nicholson BS, Sanders JE. 2003. Tissue engineering of perfused microvessels. Microvasc Res 66:59-67.

Nicosia RF, Bonanno E, Smith M,Yurchenco P. 1994a. Modulation of angiogenesis in vitro by laminin-entactin complex. Dev Biol 1641197-206.

Nicosia RF, Bonanno E, Villaschi S. 1992. Large-vessel endothelium switches to a microvascular phenotype during angiogenesis in collagen gel culture ofrat aorta. Atherosclerosis 95: 191-199.

Nicosia RF, Nicosia SV, Smith M. 1994b. Vascular endothelial growth factor, platelet-derived growth factor, and insulin-like growth factor-1 promote rat aortic angiogenesis in vitro. Am J Pathol 145:1023-1029.

Nicosia RF, Ottinetti A. 1990. Modulation of microvascular growth and morphogenesis by reconstituted basement membrane gel in

three-dimensional cultures of rat aorta: a comparative study of angiogenesis in matrigel, collagen, fibrin, and plasma clot. In Vitro Cell Dev Biol 26:119-128.

Nicosia RF, Tchao R, Leighton J . 1982. Histotypic angiogenesis in vitro: light microscopic, ultrastructural, and radioautographic studies. In Vitro 18:538-549.

Nicosia RF, Tchao R, Leighton J. 1983. Angiogenesis-dependent tumor spread in reinforced fibrin clot culture. Cancer Res 43121592166.

Nicosia RF, Tuszynski GP 1994. Matrix-bound thrombospondin promotes angiogenesis in vitro. J Cell Biol 1241183-193.

Passaniti A, Taylor RM, Pili R, GuoY, Long PV, Haney JA, Pauly RR, Grant DS, Martin GR. 1992. A simple, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor. Lab Invest 67:519-528.

Pepper MS, Montesano R, Vassalli JD, Orci L. 1991. Chondrocytes inhibit endothelial sprout formation in vitro: evidence for involvement of a transforming growth factor-beta. J Cell Physiol 1461170179.

Rosen EM, Meromsky L, Setter E, Vinter DW, Goldberg ID. 1990. Quantitation of cytokine-stimulated migration of endothelium and epithelium by a new assay using microcarrier beads. Exp Cell Res 186 :22-3 1 .

Sage EH, Vernon RB. 1994. Regulation of angiogenesis by extracellular matrix: the growth and the glue. J Hypertens Suppl 12:S145152.

Vernon RB, Angello JC, Iruela-Arispe ML, Lane TF, Sage EH. 1992. Reorganization of basement membrane matrices by cellular traction promotes the formation of cellular networks in vitro. Lab Invest 66:536-547.

Vernon RB, Gooden MD. 2002. New technologies in vitro for analysis of cell movement on or within collagen gels. Matrix Biol 211661669.

Vernon RB, Lara SL, Drake CJ, Iruela-Arispe ML, Angello JC, Little CD, Wight TN, Sage EH. 1995. Organized type I collagen influences endothelial patterns during “spontaneous angiogenesis in vitro”: planar cultures as models of vascular development. In Vitro Cell Dev Biol Anim 31:120-131.

Vernon RB, Sage EH. 1999. A novel, quantitative model for study of endothelial cell migration and sprout formation within three-dimensional collagen matrices. Microvasc Res 57:118-133.

Villaschi S, Nicosia RF. 1993. Angiogenic role of endogenous basic fibroblast growth factor released by rat aorta after injury. Am J Pathol 1431181-190.

Chrobak KM, Potter DR, Tien J , “Formation of perfused, functional microvascular tubes in vitro”, Microvascular Research 71 (May 2006), 185-196.

Neumann, Thomas, 2005, Grant Abstract, Grant No. 1 R21 HL081152-01 awarded by NIH National Heart, Lung, and Blood Institute.

Ratner, Buddy D., 2003, Grant Abstract, Grant No. 5R24HL0643 8704 awarded by NIH National Heart, Lung, and Blood Institute. Nicosia, R.F., Ottinetti, A., Growth of Microvessels in Serum-Free Matrix Culture of Rat Aorta, Laboratory Investigation, vol. 63, No. 1, p. 115-122, 1990.

L’Heureux, N. et al., A completely biological tissue-engineered human blood vessel, The FASEB Journal, vol. 12, January, p. 47-56, 1998.

Maniotis, A. J . et al., Vascular Channel Formation by Human Melanoma Cells in Vivo and in Vitro: Vasculogenic Mimicry, American Journal of Pathology, vol. 155, No. 3, p. 739-752, Sep. 1999. Neumann, T. et al., Tissue Engineering of Skeletal Muscle Using Polymer Fiber Arrays, Tissue Engineering, vol. 9, No. 5, p. 995-1003, 2003.

Lin, A. Y. et al., Distinguishing Fibrovascular Septa From Vasculogenic Mimicry Patterns, Arch. Pathol. Lab. Med., vol. 129, p. 884-892, Jul. 2005.