New born child tissue engineering bladder piece and preparation method thereof

A tissue engineering, neonatal technology, applied in the direction of bone/connective tissue cells, artificial cell constructs, medical science, etc., can solve the problems of unsuitable bladder tumor patients, cell aging, death and other problems, achieve good cell adhesion ability, The effect of obvious tissue thickening and rapid cell proliferation

Inactive Publication Date: 2011-07-13
SHANGHAI CHILDRENS HOSPITAL
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, autologous bladder transitional epithelial and smooth muscle cells have disadvantages as seed cells: ① When the body’s organs are in terminal failure, tissue samples for expanding sufficient cells may not be available, and as differentiated and mature cells, when cultured in vitro, Not only is the proliferation rate slow, but there are also problems such as dedifferentiation; ②Due to the shortening of the continuity of telomeres, normal somatic cells can only undergo limited division and passage. If the limit of passage is reached, the cells will age and die; ③Neurogenic The smooth muscle cells of normal bladder are different from normal cells in terms of cell proliferation, contractility and cell adhesion; ④ It is not suitable for patients with bladder tumors
Embryonic stem cells have limitations in their application due to issues such as ethics, sources, immune rejection, and susceptibility to tumorigenesis under unsuitable conditions, while adult stem cells also have defects such as cell number and immune rejection, and it is also difficult to obtain materials for newborns

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
  • New born child tissue engineering bladder piece and preparation method thereof
  • New born child tissue engineering bladder piece and preparation method thereof
  • New born child tissue engineering bladder piece and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] 1. Isolation, sorting and induction culture of hAFCSCs:

[0036] Amniotic fluid samples come from pregnant women undergoing prenatal diagnosis in the second trimester (4-6 months). Ultrasound-guided amniocentesis is used to obtain 20-40ml amniotic fluid into a 50ml sterile centrifuge tube, which is sealed and stored in a 4°C refrigerator and quickly transferred to a laminar flow sterile tube. The ultra-clean bench in the cell culture room. After filtering through a sterile 100-mesh sieve, centrifuge at 1000 rpm for 5 min at 4°C. After centrifugation, the supernatant was discarded, and 5ml of culture medium was added to the tube for cultivation. After 8-10 days, clones with good growth and uniform morphology were picked with the cloning cup to obtain hAFCSCs. After continuing to culture and expand in vitro, select the 4th passage hAFCSCs. When they grow to 70-80% confluence, replace with the basic induction medium and add induction factors to the culture dish every 24 ...

Embodiment 2

[0047] Cell fluorescent immunostaining was performed on the induced cells obtained in Example 1, and it was confirmed that hAFCSCs induced by the above induction scheme expressed smooth muscle cell marker smooth muscle actin (α-smooth muscle actin, α-SMA) ( figure 2 A), troponin (calponin, CALP) ( figure 2 B) and myosin heavy chain (MHC) ( figure 2 C), demonstrating that our induction method can successfully differentiate hAFCSCs into smooth muscle-like cells.

[0048] Immunofluorescent staining of tissue immunofluorescent staining found that smooth muscle cell marker protein α-SMA, CALP, and MHC all expressed ( image 3 arrow pointing).

[0049] The PGA material support and compound that obtain in example 1 two, three are carried out scanning electron microscope observation and visible non-woven PGA support material is irregular grid-like structure ( figure 2 A), the grid size is conducive to the penetration and migration of hAFCSCs into the scaffold; at one week, it c...

Embodiment 3

[0050] Example 3 Evaluation of the mechanical effect of the composite of hAFCSCs planted in vivo and PGA material scaffold:

[0051] Trim the sample obtained in Example 14 into a rectangular shape with a size of about 15mm×10mm×1.5mm. The specimen is fixed on a universal electronic stretching machine (instron 5969), using a 50N tension sensor at a speed of 5mm / min. Load the sample, detect the maximum tensile stress (that is, the maximum load), the maximum elongation rate, the deformation at the maximum load and other indicators, and the measured data are analyzed and processed by the statistical software SPSS.11.0, and the mechanical indicators of each group Expressed as mean ± standard deviation (Table 1). The comparison of the mechanical data between the samples in each group was performed by the Student’s t test, and P Figure 4A ); the mechanical experiment group was compared with the mechanical positive control group (P=0.0103), and the difference was significant ( Figure ...

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

No PUM Login to view more

Abstract

The invention relates to the technical field of medical tissue engineering, in particular to a new born child tissue engineering bladder piece and a preparation method thereof. The new born child tissue engineering bladder piece provided by the invention is characterized by comprising smooth muscle-like cells formed by differentiation and a degradable scaffold material; the smooth muscle-like cells wrap the degradable scaffold material and are flaky; and the smooth muscle-like cells express alpha smooth muscle actin (alpha-SMA), calponin (CALP) and major histocompatibility complex (MHC). In terms of mechanical indexes, the new born child tissue engineering bladder piece has similar mechanical tensile characteristic with normal bladder smooth muscle tissues, which helps to restore the consistency of the bladder compliance during rebuilding of filling of bladder.

Description

technical field [0001] The invention relates to the technical field of medical tissue engineering, in particular to a neonatal tissue engineering bladder tablet and a preparation method thereof. Background technique [0002] Congenital bladder malformations include bladder exstrophy, hypoplasia, duplication bladder, patent urachal, megabladder syndrome, and bladder diverticulum. For example, bladder exstrophy is due to abnormal development of the cloacal membrane during the embryonic period, resulting in congenital lower abdominal wall and Bladder front wall defect, bladder posterior wall eversion, bladder deformity with exposed mucosa, this disease is rare, about one case in 10,000 to 40,000 newborns, males are more than females, about 2 to 8:1, due to bladder exversion, bladder Exposure of the mucous membrane and ureteral orifice can easily lead to urinary incontinence and ascending nephronitis. Half of untreated patients die in childhood. Therefore, children with congeni...

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
Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38C12N5/071C12N5/0775C12N5/0789
Inventor 陈方周君梅高同斌
Owner SHANGHAI CHILDRENS HOSPITAL
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