Postpartum cells derived from umbilical cord tissue, and methods of making and using the same

a postpartum cell and umbilical cord technology, applied in the field of mammals, can solve the problems of time-consuming and expensive pre-testing, method not distinguishing the desired cells, and variation in the composition of serum lot-to-lot,

Inactive Publication Date: 2006-10-05
DEPUY SYNTHES PROD INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0033] Methods are provided of inducing the cells to differentiate along a pathway towards progenitors of various cells, or even into terminally differentiated cells themselves. Such cells have ut

Problems solved by technology

As such, the method did not distinguish the desired cells from the different cell types present in Wharton's Jelly, but rather relied on migration from the tissue or selecting growth conditions favoring prechondrocytes.
In spite of the advantages of including bovine serum (or serum of other species), there are, however, a number of disadvantages to supplementing medium with animal serum instead of using chemically-defined or serum-free media when culturing therapeutic cells or producing biologics.
Furthe

Method used

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  • Postpartum cells derived from umbilical cord tissue, and methods of making and using the same

Examples

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example 1

Isolation of Cells from Postpartum Umbilicus Tissues

[0203] Postpartum umbilicus tissues were obtained upon birth of either a full term or pre-term pregnancy. Cells were harvested from five separate donors of umbilicus tissue. Different methods of cell isolation were tested for their ability to yield cells with: 1) the potential to differentiate into cells with different phenotypes, a characteristic common to stem cells, or 2) the potential to provide critical trophic factors useful for other cells and tissues.

Methods & Materials

[0204] Umbilical cell isolation. Umbilical cords were obtained from National Disease Research Interchange (NDRI, Philadelphia, Pa.). The tissues were obtained following normal deliveries. The cell isolation protocols were performed aseptically in a laminar flow hood. To remove blood and debris, the cord was washed in phosphate buffered saline (PBS; Invitrogen, Carlsbad, Calif.) in the presence of penicillin at 100 Units / milliliter and streptomycin at 100 ...

example 2

Growth Characteristics of Umbilicus-Derived Cells

[0222] The cell expansion potential of umbilicus-derived cells was compared to other populations of isolated stem cells. The process of cell expansion to senescence is referred to as Hayflick's limit (Hayflick L. The longevity of cultured human cells. J. Am. Geriatr. Soc. 22(1):1-12, 1974; Hayflick L. The strategy of senescence. Gerontologist 14(1):37-45), 1974).

Materials and Methods

[0223] Gelatin-coating flasks. Tissue culture plastic flasks were coated by adding 20 milliliters 2% (w / v) gelatin (Type B: 225 Bloom; Sigma, St Louis, Mo.) to a T75 flask (Corning Inc., Corning, N.Y.) for 20 minutes at room temperature. After removing the gelatin solution, 10 milliliters phosphate-buffered saline (PBS) (Invitrogen, Carlsbad, Calif.) were added and then aspirated.

[0224] Comparison of expansion potential of umbilicus-derived cells with other cell populations. For comparison of growth expansion potential the following cell populations w...

example 3

Growth of Umbilicus-Derived Cells in Medium Containing D-Valine

[0241] It has been reported that medium containing D-valine instead of the normal L-valine isoform can be used to selectively inhibit the growth of fibroblast-like cells in culture (Hongpaisan, 2000; Sordillo et al., 1988). Experiments were performed to determine whether umbilicus-derived cells could grow in medium containing D-valine.

Methods & Materials

[0242] Umbilicus-derived cells (P5) and fibroblasts (P9) were seeded at 5,000 cells / cm2 in gelatin-coated T75 flasks (Corning, Corning, N.Y.). After 24 hours the medium was removed and the cells were washed with phosphate buffered saline (PBS) (Gibco, Carlsbad, Calif.) to remove residual medium. The medium was replaced with a modified Growth Medium (DMEM with D-valine (special order Gibco), 15% (v / v) dialyzed fetal bovine serum (Hyclone, Logan, Utah), 0.001% (v / v) betamercaptoethanol (Sigma), penicillin at 50 Units / milliliter and streptomycin at 50 milligrams / millilit...

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Abstract

Cells derived from human umbilical cords are disclosed along with methods for their therapeutic use. Isolation techniques, culture methods and detailed characterization of the cells with respect to their cell surface markers, gene expression, and their secretion of trophic factors are described.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This is a continuation-in-part of U.S. patent application Ser. No. 10 / 877,012, filed Jun. 25, 2004, which claims benefit of U.S. Provisional Application Ser. No. 60 / 483,264, filed Jun. 27, 2003, the entire contents of each of which are incorporated by reference herein. This also claims benefit of U.S. Provisional Application No. 60 / 639,088, filed Dec. 23, 2004, the entirety of which is also herein incorporated by reference.FIELD OF THE INVENTION [0002] This invention relates to mammalian, preferably human, cell therapy, and more particularly to isolated cells derived from postpartum umbilicus, methods of deriving such cells, and methods of their use and growth on various media, including media which are serum-containing or serum-free. BACKGROUND OF THE INVENTION [0003] As the modern understanding of disease has advanced, the potential utility of cell therapy for improving the prognosis of those afflicted has resulted in increased intere...

Claims

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Application Information

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IPC IPC(8): C12N5/00C12N9/00C12N9/26C12N5/074C12N5/0793
CPCC12N5/0607C12N5/0619C12N2500/32C12N2500/99C12N2506/03C12N2501/115C12N2501/119C12N2501/135C12N2501/11A61P1/16A61P1/18A61P7/02A61P9/00A61P13/12A61P19/00A61P25/00A61P27/02A61P29/00A61P37/02A61P37/06A61P43/00C12N2500/90A61K35/50A61K35/51
Inventor HARRIS, IANMESSINA, DARINKIHM, ANTHONYSEYDA, AGNIESZKACOLTER, DAVID
Owner DEPUY SYNTHES PROD INC
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