Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof

Inactive Publication Date: 2005-12-22
FURCHT LEO +2
View PDF0 Cites 90 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0047] In another preferred embodiment, the

Problems solved by technology

However, there are at least two major problems associated with organ and tissue transplantation.
First, there is a shortage of donor organs and tissues.
The second major problem is the potential incompatibility of the transplanted tissue with the immune system of the recipient.
Because the donated organ or tissue is recognized by the host immune system as foreign, immunosuppressive medications must be provided to the patient at a significant cost-both financially and physically.
Unfortunately, xenotransplantation does not overcome the problem of tissue incompatibility, but instead exacerbates it.
Furthermore, according to the Centers for Disease Control, there is evidence that damaging viruses cross species barriers.
Stem cells which differentiate only to form cells of hematopoietic lineage, however, are unable to provide a source of cells for repair of other damaged tissues, for example, heart or lung tissue damaged by high-dose chemotherapeutic agents.
The most commonly used source of NSC is allogeneic fetal brain, which poses both immunological and ethical problems.
As it is not known whether pre-existing neural pathology will affect the ability of NSC to be cultured and induced to differentiate into neuronal and glial cells ex vivo, and because additional surgery in an already diseased brain may aggravate the underlying disease, this approach is less attractive.
This cell is capable of differentiating to f

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof
  • Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof
  • Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

Selection. Culture and Characterization of Mouse Multipotent Adult Stem Cells (mMASC)

Cell Isolation and Expansion

[0093] All tissues were obtained according to guidelines from the University of Minnesota IACUC. BM mononuclear cells (BMMNC) were obtained by ficoll-hypaque separation of BM was obtained from 5-6 week old ROSA26 mice or C57 / BL6 mice. Alternatively, muscle and brain tissue was obtained from 3-day old 129 mice. Muscles from the proximal parts of fore and hind limbs were excised from and thoroughly minced. The tissue was treated with 0.2% collagenase (Sigma Chemical Co, St Louis, Mo.) for 1 hour at 37° C., followed by 0.1% trypsin (Invitrogen, Grand Island, N.Y.) for 45 minutes. Cells were then triturated vigorously and passed through a 70-urn filter. Cell suspensions were collected and centrifuged for 10 minutes at 1600 rpm. Brain tissues was dissected and minced thoroughly. Cells were dissociated by incubation with 0.1% trypsin and 0.1% DNAse (Sigma) for 30 minutes at ...

example 2

Selection and Culture of Rat Multipotent Adult Stem Cells (rMASC)

[0103] BM and MNC from Sprague Dawley or Wistar rats were obtained and plated under conditions similar for mMASC. After 21-28 days, cells were depleted of CD45+ cells, and the resulting CD45 cells were subcultured at 10 cells / well.

[0104] Similar to mMASC, rMASC have been culture expanded for >100 PDs. Expansion conditions of rat MASC culture required the addition of EGF, PDGF-BB and LIF and culture on FN, but not collagen type 1, laminin or Matrigel™. rMASC were CD44, CD45 and MHC class I and II negative, and expressed high levels of telomerase. The ability of a normal cell to grow over 100 cell doublings is unprecedented, unexpected and goes against conventional dogma of more than two decades.

[0105] Rat MASC that had undergone 42 PDs, 72 PDs, 80 PDs, and 100 PDs, were harvested and telomere lengths evaluated. Telomeres did not shorten in culture, as was determined by Southern blot analysis after 42 PDs, 72 PDs, 80 ...

example 3

Selection and Culture of Human Multipotent Adult Stem Cells (hMASC)

[0106] BM was obtained from healthy volunteer donors (age 2-50 years) after informed consent using guidelines from the University of Minnesota Committee on the use of Human Subject in Research. BMMNC were obtained by Ficoll-Paque density gradient centrifugation and depleted of CD45+ and glycophorin-A+ cells using micromagnetic beads (Miltenyii Biotec, Sunnyvale, Calif.).

[0107] Expansion conditions: 5×103 CD45− / GlyA− cells were diluted in 200 μL expansion medium [58% DMEM-LG, 40% MCDB-201 (Sigma Chemical Co, St Louis, Mo.), supplemented with 1× insulin-transferrin-selenium (ITS), 1×-linoleic-acid bovine serum albumin (LA-BSA), 10−8 M Dexamethasone, 10−4 M ascorbic acid 2-phosphate (all from Sigma), 100 U penicillin and 1,000 U streptomycin (Gibco)] and 0-10% fetal calf serum (FCS) (Hyclone Laboratories, Logan, Utah) with 10 ng / ml of EGF (Sigma) and 10 ng / ml PDGF-BB (R&D Systems, Minneapolis, Minn.)] and plated in we...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
Fractionaaaaaaaaaa
Timeaaaaaaaaaa
Timeaaaaaaaaaa
Login to view more

Abstract

Methods and compositions are provided for circularizing target sequences in a sample. In particular, ligation oligonucleotides are employed to selectively hybridize with the target such that the target can be ligated into a closed circular target. Rolling circle amplification can then be performed directly on the target sequence for subsequent detection and analysis.

Description

RELATED CASES [0001] This application claims the benefit of U.S. Provisional Application No. 60 / 343,386, filed Oct. 25, 2001, U.S. Provisional Application No. 60 / 310,625, filed Aug. 7, 2001, U.S. Provisional Application No. 60 / 269,062, filed Feb. 15, 2001, U.S. Provisional Application No. 60 / 268,786, filed Feb. 14, 2001, which are hereby incorporated by reference for all purposes. Applicants also claim priority of WO 01 / 11011, 60 / 147,324 and 60 / 164,650 and these applications are hereby incorporated by reference into this text; any teachings therein may be used in the practice of this invention. The present application is a continuation-in-part of 15′ WO 01 / 11011, which is attached herein at Appendix 1 and is part of the present application. Documents incorporated by reference into this text are not admitted to be prior art.FIELD OF THE INVENTION [0002] The present invention relates generally to mammalian multipotent adult stem cells (MASC), and more specifically to methods for obtai...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): A01K67/027A01N63/00A61D19/04A61K35/12A61K35/14A61K35/28A61K35/30A61K35/32A61K35/39A61K35/407A61K35/44A61K39/00A61K45/00A61K47/46A61K48/00A61L27/00A61L31/00A61P1/00A61P1/16A61P1/18A61P3/00A61P3/10A61P7/00A61P7/04A61P7/06A61P7/08A61P9/00A61P9/10A61P13/10A61P13/12A61P15/00A61P17/00A61P19/04A61P19/08A61P21/00A61P25/00A61P25/02A61P27/02A61P31/00A61P31/04A61P31/10A61P31/12A61P35/00A61P37/00A61P37/06A61P39/02A61P41/00A61P43/00C07K1/00C07K14/00C12NC12N5/00C12N5/02C12N5/071C12N5/074C12N5/079C12N5/0793C12N15/00C12N15/85C12N15/873C12Q1/02C12Q1/68G01F19/00G01N33/48G01N33/50
CPCA01K67/0271C12N2517/02A01K2217/075A01K2227/105A01K2227/106A61K35/12A61K39/001A61K48/00C12N5/0607C12N5/0619C12N5/0622C12N5/067C12N15/873C12N2501/11C12N2501/113C12N2501/115C12N2501/117C12N2501/119C12N2501/12C12N2501/135C12N2501/235C12N2501/237C12N2502/08C12N2502/30C12N2503/00C12N2506/03A01K2217/05A61P1/00A61P1/16A61P1/18A61P13/10A61P13/12A61P15/00A61P17/00A61P19/04A61P19/08A61P21/00A61P25/00A61P25/02A61P27/02A61P3/00A61P31/00A61P31/04A61P31/10A61P31/12A61P35/00A61P37/00A61P37/06A61P39/02A61P41/00A61P43/00A61P7/00A61P7/04A61P7/06A61P7/08A61P9/00A61P9/10A61P3/10
Inventor FURCHT, LEOVERFAILLIE, CATHERINEREYES, MORAYMA
Owner FURCHT LEO
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap