Pluripotent human stem cell technology, method and product of small molecule oriented tissue and organ regeneration

A technology of stem cells and human cells, applied in the field of biomedicine, can solve problems such as laborious purification or separation steps, unsuitable for commercial and clinical applications, unsuitable for clinical applications or human trials

Inactive Publication Date: 2016-11-23
苏州博瑞斯坦生物医药有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purification or isolation steps of those conventional pluripotent human stem cell differentiation methods are laborious, expensive, and time-consuming, yield only a small amount of desired cells, and are not suitable for commercial and clinical applications
More and more scientific evidence shows that those traditional differentiation methods lead to inefficiency, instability, incomplete differentiation, poor performance, and high tumor risk after transplantation of cell derivatives and tissue engineering constructs
According to current methods used in the field, pluripotent human ste

Method used

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  • Pluripotent human stem cell technology, method and product of small molecule oriented tissue and organ regeneration
  • Pluripotent human stem cell technology, method and product of small molecule oriented tissue and organ regeneration
  • Pluripotent human stem cell technology, method and product of small molecule oriented tissue and organ regeneration

Examples

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Effect test

Embodiment 1

[0122] This example provides a method to induce high-quality clinical-grade xeno-free pluripotent human stem cells that are mass-manufactured (cGMP) by small molecule directed differentiation into functional human nerves or cells that can be used for tissue engineering, Drug development, drug testing, and cell therapy.

[0123] Undifferentiated pluripotent human embryonic stem cells were seeded and maintained in a defined culture system containing DMEM / F-12 or KO-DMEM (knockout-DMEM) (80%), knockout serum replacement (KO) (20%) , L-alanyl-L-glutamine or L-GLN (2mM), MEM non-essential amino acids (MNAA, 1X), mercaptoethanol (0.1mM), purified human laminin (40 μg / ml) [ Dilute 1:30 in cold DMEM / F-12] or laminin / collagen as matrix protein, and bFGF (basic fibroblast growth factor, 20 nanograms / milliliter [ng / ml]). Knockout serum substitutes (KO) can be replaced with defined human elements containing MEM essential amino acids (MEAA, 1X), human insulin (20 μg / ml) and ascorbic acid ...

Embodiment 2

[0125] This example provides a method for the maintenance of high-quality clinical-grade xeno-free direct from large-scale manufacturing (cGMP) using current good manufacturing practice (cGMP) in a defined culture system through the use of retinoic acid (RA)-induced Directed and efficient differentiation of human embryonic stem cells into neuroectoderm by promoting the nuclear translocation of the neuron-specific transcription factor Nurr-1, and triggering the high-efficiency, high-purity, and large-scale development of lineage-specific neurons into human neural precursor cells and human Neuronal cells (>90%) ( figure 1 ,3,5).

[0126] Neuronal lineage-specific differentiation directly from the pluripotent state of human embryonic stem cells. Inoculate and maintain undifferentiated high-quality clinical-grade xeno-free pluripotent human embryonic stem cells in a defined culture system for 3 days, add RA (0.01mM) and continue to culture for 4-5 days under defined culture condi...

Embodiment 3

[0130] This example provides a method to induce xeno-free human cultures maintained in a defined culture system directly from high-quality clinical-grade mass-manufactured (cGMP) using current good manufacturing practice (cGMP) using nicotinamide (NAM). Directed and efficient differentiation of cardiac mesoderm from embryonic stem cells promotes the expression of the early cardiomyocyte-specific transcription factor CSX / Nkx2.5 and triggers the efficient, high-purity, and large-scale development of cardiac lineage-specific human cardiac precursors and beating human myocardium cells (>90%) ( figure 2 ,4,6).

[0131]Cardiomyocyte lineage-specific differentiation of pluripotent human embryonic stem cells using small molecules. Inoculate and maintain undifferentiated high-quality clinical-grade xeno-free pluripotent human embryonic stem cells in a defined culture system for 3 days, add NAM (10mM) and continue to culture for 4-5 days under defined culture conditions. These cardia...

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Abstract

The invention provides pluripotent human stem cell technology, method and product of small molecule oriented tissue and organ regeneration. The technology provided in the invention can directly and high-effectively convert pluripotent human stem cells into high-purity human cells in specific pedigree in large scale, which are high in quality, are clinical-grade and have no foreign matters, through small molecule oriented differentiation induction. The invention provides an approach of producing or manufacturing the pluripotent human stem cells, which are high in quality, are clinical-grade and have no foreign matters, and a functional human cell therapy derivative thereof in large scale for application of cell therapy and commercialization. In particular, the invention creatively provides a new commercial source of high-purity functional human neurons and myocardial cells for medical treatment of repairing central nervous system or cardiac muscle.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular, the invention relates to the pluripotent human stem cell technology, method and product for small molecule directed tissue and organ regeneration. Background technique [0002] Undifferentiated human embryonic stem cells (human embryonic stem cells [hESC]) have long-term stable culture growth and unlimited intrinsic potential to differentiate into various types of human cells. Human embryonic stem cells, derived from the inner cell mass of the blastocyst, the blastocyst, or the ectoderm, not only provide a powerful in vitro model system for studying human embryonic development, but also provide a large supply of human cells for disease-specific lineages. provides a versatile repository of in vitro derivatives [1-3]. However, how to effectively and predictably direct the broad differentiation potential of pluripotent human stem cells to generate desired human cell types has been a major ...

Claims

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Application Information

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IPC IPC(8): C12N5/079C12N5/071A61L27/38
Inventor 黄雪珺·帕森斯
Owner 苏州博瑞斯坦生物医药有限公司
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