Chemistry: molecular biology and microbiology – Animal cell – per se ; composition thereof; process of... – Solid support and method of culturing cells on said solid...
Reexamination Certificate
2000-11-09
2004-08-17
Lankford, Jr., Leon B. (Department: 1651)
Chemistry: molecular biology and microbiology
Animal cell, per se ; composition thereof; process of...
Solid support and method of culturing cells on said solid...
C435S283100, C435S305100, C435S325000, C435S402000
Reexamination Certificate
active
06777234
ABSTRACT:
TECHNICAL FIELD
This invention relates to the field of tissue engineering, and more particularly to a mammalian muscle construct and a method for producing the construct in vitro.
BACKGROUND ART
At present, three-dimensional tissues are capable of being produced in vitro using various types of cells. For example, U.S. Pat. No. 5,443,950 issued to Naughton et al. describes three-dimensional cultures for bone marrow, skin, liver, vascular, and pancreatic tissues which are grown within synthetic matrices. In these tissues as well as others, investigators have been successful in proliferating cells and tissues in vitro such that the resulting three-dimensional tissues, termed “organoids” or “constructs”, display many of the characteristics of their in vivo counterparts. These constructs have a variety of foreseeable applications, ranging from transplantation in vivo to functional and pharmacological testing in vitro.
In terms of muscle tissue, in vitro constructs of smooth muscle, cardiac muscle, and skeletal muscle have each been formulated. For example, U.S. Pat. No. 5,618,718 issued to Auger et al. describes the production of a contractile smooth muscle cell construct, and U.S. Pat. No. 4,605,623 issued to Malette et al. describes a method for cultivating the three-dimensional growth of cardiac myocytes. These smooth muscle and cardiac muscle constructs were each developed using mammalian muscle cells, specifically, human muscle cells.
In contrast, the majority of skeletal muscle organoids have been developed using avian muscle cells. In particular, a series of studies conducted by Vandenburgh and colleagues involved the production of organoids from avian muscle cells grown on an expandable, SILASTIC® membrane (Vandenburgh,
In Vitro Cell. Dev. Biol
. 24: 609-619, 1988; Vandenburgh et al.,
Am. J. Physiol
. 256 (
Cell Physiol
. 25): C674-C682, 1989; Vandenburgh et al.,
In Vitro Cell. Dev. Biol
. 25: 607-619, 1989; Vandenburgh et al.,
FASEB J
. 5: 2860-2867, 1991). Since avian muscle is structurally and functionally distinct from mammalian muscle, organoids developed from avian muscle have no direct clinical application. A few skeletal muscle constructs have been developed using mammalian muscle grown within a synthetic matrix (Vandenburgh et al.,
Hum. Gene Ther
. 7: 2195-2200, 1996; Shansky et al.,
In Vitro Cell Dev. Biol
. 33: 659-661, 1997). However, the constructs in these studies originated from cells extracted from neonatal rats or immortal cell lines (C2C12) established from C3H mice which, due to their age or pathology, have limited clinical significance.
Previous methods of organoid production have additional drawbacks. First, in the majority of the studies by Vandenburgh and colleagues described above, as well as in U.S. Pat. Nos. 4,940,853 and 5,153,136, both issued to Vandenburgh, mechanical strain is applied to the skeletal muscle organoids for their proper development, such that complex mechanical fixturing and control electronics are required. Second, both the mammalian and avian skeletal muscle constructs have a limited in vitro life span of approximately four weeks, preventing their use for long-term functional or pharmacological studies.
Perhaps the most serious drawback of previous studies involving the growth of three-dimensional tissues is that the type of anchor systems to which the tissues attach restricts the ability to functionally evaluate the tissues. For instance, when a synthetic membrane or matrix is utilized, the contractile function of the organoids may be difficult to determine separate from the matrix material due to the mechanical preloads of the matrix material. When synthetic anchors such as stainless steel pins or mesh are employed, the tissue merely grows around the anchors instead of into them, such that there is a large discontinuity in mechanical impedance. This discontinuity creates a stress concentration, which could lead to cell damage when the tissue contracts.
DISCLOSURE OF THE INVENTION
Therefore, it is an object of the present invention to provide a mammalian muscle construct which is developed in vitro from cells extracted from mammals of any age.
It is another object of the present invention to provide an anchor system for forming a mammalian muscle construct wherein the anchor system does not restrict the ability to functionally evaluate the construct.
It is a further object of the present invention to provide a non-synthetic anchor system for forming a mammalian muscle construct.
It is a further object of the present invention to provide a method for producing mammalian muscle constructs which does not require the application of external mechanical strain.
It is still another object of the present invention to provide a mammalian muscle construct which is capable of being maintained in vitro for longer than four weeks.
Accordingly, a mammalian muscle construct and a method for producing the construct are provided. The mammalian muscle construct includes a substrate and a plurality of separate anchors secured to the substrate. Myogenic precursor cells are provided on the substrate with at least some of the cells in contact with the anchors. The myogenic precursor cells are cultured in vitro under conditions to allow the cells to become confluent between the anchors. The anchors are receptive to the cells and allow the cells to attach thereto, such that placement of the anchors controls the size and shape of the muscle construct formed.
For use in producing the mammalian muscle construct, an anchor system for controllably forming tissue from precursor cells in vitro and a method for making the anchor system are provided. The anchor system includes a substrate and a plurality of separate fragments of biocompatible material secured to the substrate. Cell adhesion molecules are associated with each fragment to facilitate attachment of the precursor cells to the fragment. Therefore, the placement of the fragments on the substrate defines an area for confluence of the cells to control the size and shape of the tissue formed.
The above objects and other objects, features, and advantages of the present invention are more readily understood from a review of the attached drawings and the accompanying specification and claims.
REFERENCES:
patent: 4605623 (1986-08-01), Malette et al.
patent: 4940853 (1990-07-01), Vandenburgh
patent: 5153136 (1992-10-01), Vandenburgh
patent: 5443950 (1995-08-01), Naughton et al.
patent: 5618718 (1997-04-01), Auger et al.
patent: 5756350 (1998-05-01), Lee et al.
patent: 6207451 (2001-03-01), Dennis et al.
Herman A. Vandenburgh Et Al., Skeletal Muscle Growth is Stimulated by Intermittent Stretch-Relaction In Tissue Culture, The American Physiological Society, 1989, pp. C674-C682.
Herman A. Vandenburgh, A Computerized Mechanical Cell Stimulator for Tissue Culture: Effects on Skeletal Muscle Organogenesis, In Vitro Cellular & Developmental Biology, vol. 24, No. 7, Jul., 1988, pp. 609-619.
Herman A. Vandenburgh Et Al., Longitudinal Growth of Skeletal Myotubes in Vitro in a New Horizontal Mechanical Cell Stimulator, In Vitro Cellular & Developmental Biology, vol. 25, No. 7, Jul., 1989, pp. 607-616.
Shansky Et Al., Letter to the Editor: A Simplified Method for Tissue Engineering Skeletal Muscle Organoids In Vitro, In Vitro Cell. Dev. Biol., Oct., 1997, pp. 659-661.
Herman A. Vandenburgh Et Al., Computer-Aided Mechanogenesis of Skeletal Muscle Organs from Single Cells in Vitro, The FASEB Journal, vol. 5, Oct. 1991, pp. 2860-2867.
Herman A. Vandenburgh Et Al., Brief Report: Tissue-Engineered Skeletal Muscle Organoids for Reversible Gene Therapy, Human Gene Therapy (Nov. 10, 1996), pp. 2195-2200.
Dennis Robert G.
Faulkner John A.
Kosnik Paul
Kuzon, Jr. William M.
Brooks & Kushman P.C.
Lankford , Jr. Leon B.
The Regents of the University of Michigan
LandOfFree
Mammalian muscle construct and method for producing same does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Mammalian muscle construct and method for producing same, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Mammalian muscle construct and method for producing same will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3313778