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Implantable construct, methods of manufacturing, and uses thereof

A technology of constructs and microcarriers, applied in general culture methods, biochemical equipment and methods, prostheses, etc., can solve problems such as harmful effects of MSCs

Pending Publication Date: 2021-09-10
AGENCY FOR SCI TECH & RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such harvesting procedures have detrimental effects on MSCs, and MSCs thus obtained often require time to recover from the harvesting procedure before regaining their full functional potential

Method used

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  • Implantable construct, methods of manufacturing, and uses thereof
  • Implantable construct, methods of manufacturing, and uses thereof
  • Implantable construct, methods of manufacturing, and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0144] Example 1 - Evaluation of key parameters for efficient chondrogenic differentiation of heMSC-LPCL microcarrier constructs

[0145] Schematic of the experimental design - Phase 1: at day 3 (early logarithmic growth with 21% cell confluency), day 5 (mid logarithmic growth with 53% cell confluency) or day 7 (with 73% cell confluency confluent heMSCs attached to LPCL microcarriers were seeded as chondrogenic heMSC-microcarrier constructs using different cell numbers per construct. Critical cell confluency and cell numbers per construct were then identified by assessing cell growth and differentiation yield at day 21. Phase 2: Evaluation of the effect of compaction of the heMSC-microcarrier constructs generated under the critical limiting conditions established in Phase 1. heMSCs were seeded with (i) centrifugation or no centrifugation and (ii) agitation or no agitation. The effect of centrifugation and / or agitation was determined by assessing cell growth and differentia...

Embodiment 2

[0174] Example 2 - hMSC-covered LPCL for allogeneic bone regeneration MC construct

[0175] This example describes the development of a combined stem cell-biomaterial therapeutic product in the form of a critically defined 50% hMSC covered LPCL MC construct that is scalable and biograftable for allogeneic bone regeneration .

[0176] Materials and methods

[0177]PCL (average Mn 45 kDa, Cat. No. 704105) and poly-L-lysine hydrobromide (PLL) (MW 70-150 kDa, Cat. No. P6282) were from Sigma-Aldrich. Fibronectin was purchased from Biological Industries. All chemical reagents were obtained from Sigma-Aldrich, and all media and supplements were purchased from ThermoFisher Scientific.

[0178] Manufacture of PCL MC

[0179] Porous PCL MCs were fabricated using a two-phase flow microfluidic device as previously reported. Briefly, PCL droplets were collected in glass graduated cylinders containing 70%–95% ethanol, and immersion in ethanol resulted in solidification of PCL dro...

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Abstract

The present invention refers to a method of manufacturing an implantable construct comprising chondrogenically differentiated cells and one or more polycaprolactone (PCL) microcarriers, an implantable construct produced using said method, and uses of the implantable construct. The present invention also refers to a method of manufacturing an implantable construct comprising mesenchymal stromal cells and one or more polycaprolactone (PCL) microcarriers, an implantable construct produced using said method, and uses of the implantable construct. The present invention further refers to a method of treating a disease or disorder associated with cartilage and / or bone defect, the method comprises administering one or more cell-free polycaprolactone (PCL) microcarriers in a patient suffering from the disease or disorder.

Description

[0001] Cross References to Related Applications [0002] This application claims priority from Singapore Application No. 10201809364P filed on 23 October 2018, the contents of which are hereby incorporated by reference in their entirety for all purposes. technical field [0003] The invention relates to the fields of cell biology, molecular biology and biotechnology. More particularly, the invention relates to the cultivation of stem cells on microcarriers. Background technique [0004] Broadly, cartilage and bone disorders describe a group of disorders characterized by degeneration or metabolic abnormalities of connective tissue, manifested by pain, stiffness, and limited movement of the affected body part. The source of these diseases can be pathological or even the result of trauma or injury. [0005] Because mature chondrocytes and osteocytes have little replicative potential, mature cartilage and bone have only a limited capacity for self-repair. For this reason, car...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077A61L27/38A61L27/18A61L27/54A61L27/58A61K35/32A61P19/02
CPCA61P19/02A61L27/46A61L27/48A61L27/3817A61L27/3852A61L2430/06A61K38/00A61K35/32C08L67/04A61L27/3612A61L27/3654C12N5/0075C12N5/0655C12N2506/1353C12N2531/00C12N2533/30
Inventor 林佑珊胡家荣S·鲁文尼W·R·N·伯奇黎坚林天麟A·舍卡兰
Owner AGENCY FOR SCI TECH & RES