Large-scale automated hollow fiber bioreactor expansion of human umbilical cord-derived mesenchymal stromal cells for neurological disorders. Neurodegenerative diseases make up a large group of neurological diseases and remain one of the greatest challenges and burdens for humanity. Diseases such as amyotrophic lateral sclerosis, Alzheimer’s disease, stroke or spinal cord injury are often associated with astrogliosis (astrogliosis) with symptoms of inflammation.
- The regenerative, paracrine and immunomodulatory properties of human mesenchymal stromal cells (hMSCs) can affect the above components, thus opening up new therapeutic possibilities for regenerative medicine.
- Of particular interest are hMSCs derived from umbilical cord (UC) tissue due to their origin, properties and lack of ethical paradigms. The aim of this study was to establish standard protocols for good manufacturing practice ( GMP ) for the operational and large-scale isolation, characterization, expansion and comparison of the properties of UC-hMSC cells collected in T vials compared to use of a large-scale bioreactor system. .
- Human UC-hMSC, isolated by culture technique from Wharton’s Jelly explant, was collected at 75% confluence and cultured in tissue culture bottles. UC-hMSCs obtained before / after cryopreservation and after collection in the bioreactor were fully characterized in terms of immunomodulatory “mesenchymicity”, oncogenicity and genetic stability, aging and cell doubling properties and gene expression traits . Our study demonstrates an efficient and simple technique for increasing UC-hMSC.
- Collecting UC-hMSCs using classical and large-scale methods did not alter UC-hMSC aging, genetic stability or carcinogenicity in vitro. We observed comparable growth and immunomodulatory capacity of fresh, frozen and expanded UC-hMSC.
- We found no differences in the ability to differentiate towards adipogenic, osteogenic, and chondrogenic lineages between classical and large-scale UC-hMSC expansion methods. Both methods made it possible to derive genetically stable cells with typical mesenchymal characteristics.
- Interestingly, we found that UC-hMSC grown in a bioreactor showed significantly increased levels of neural growth factor (NGF) mRNA expression and reduced levels of insulin growth factor (IGF), while IL4 , IL6, IL8,
- TGFb and VEGF expression levels remained at a similar level. Cultivation of UC-hMSC using a large-scale automated closed bioreactor expansion system under GMP conditions does not alter basic “mesenchymal” characteristics and cell quality.
- Our study was designed to pave the way for the translation of known basic research data on human UC-MSC into future clinical trials in patients with neurological and immunocompromised disorders.
- Industrial production of UC-hMSC will then be approved by regulatory authorities under the criteria for advanced therapy medicinal products (ATMP) prior to clinical use and approval for use in patients.
The relaxation factor of endothelial origin produced and released by the arteries and veins is nitric oxide.
- The aim of this study was to determine whether nitric oxide (NO) is responsible for vascular smooth muscle relaxation induced by endothelial-derived relaxation factor (EDRF). EDRF is an unstable humoral substance released by arteries and veins that mediates the action of dependent endothelial vasodilators.
- NO is an unstable, endothelium-independent vasodilator that is released by vasodilators such as nitroprusside and glyceryl trinitrate. We have repeatedly observed that the effect of NO on vascular smooth muscle is very similar to that of EDRF. In the present study, the vascular effect of EDRF released from a pulmonary artery and a perfused vein of cattle was compared with that of NO delivered by overflow through endothelial strips of cascading arteries and veins.
- EDRF was indistinguishable from NO as both were unstable (t1 / 2 = 3-5 s), inactivated by pyrogallol or superoxide anion, stabilized by superoxide dismutase, and inhibited by oxyhemoglobin or potassium. Both EDRF and NO produced a comparable increase in the cyclic accumulation of GMP in the artery and vein, and this cyclic accumulation of GMP was inhibited by pyrogallol, oxyhemoglobin, potassium and methylene blue.
- EDRF was chemically identified as NO or a labile nitrous species by two procedures. First, like NO, EDRF released from newly isolated aortic endothelial cells reacted with hemoglobin to form nitrosylemoglobin.
- Second, EDRF and NO similarly promoted the diazotization of sulphanilic acid and produced the same reaction product after coupling with N- (1-naphthyl) ethylenediamine. Therefore, EDRF released from an artery and a vein has biological and chemical properties identical to NO.
The cyclic GMP -AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway.
The presence of DNA in the cytoplasm of mammalian cells is a dangerous signal that triggers a host immune response, such as the production of type I interferons. Cytosolic DNA induces interferons by producing cyclic guanosine-adenosine monophosphate monophosphate (cyclic GMP -AMP or cGAMP) which binds to the STING adapter protein and activates them.
Through biochemical fractionation and quantitative mass spectrometry, we identified cGAMP synthase (cGAS), which belongs to the nucleotidyltransferase family. The overexpression of cGAS activated the transcription factor IRF3 and induced interferon-β in a STING-dependent manner.
The impact of cGAS inhibited the activation of IRF3 and the induction of interferon-β by DNA transfection or DNA virus infection. cGAS binds to DNA in the cytoplasm and catalyzes the synthesis of cGAMP. These results indicate that cGAS is a cytosolic DNA sensor that induces interferons by producing the second messenger cGAMP.
Providing epidemiological, virological and immunological parameters in HIV + patients defined as long-term progression (LTnP) may be the key to understanding the natural history of HIV disease and its therapeutic approach.
- The aim of this study was to identify factors that delay the onset of HIV-related symptoms in HIV-infected patients with low progression. We interviewed several people defined as LTnP according to the following criteria.
- Patients were asymptomatic with documented HIV infection for at least 8 years, had never been treated with antiretroviral therapy and had CD4 levels always above 500 / cmm. They were tested every 6 months for the following tests: virus isolation, viral strain characterization; Quantitative DNA-PCR; HIV p24 antigen in serum, plasma viraemia, neutralizing antibodies to autologous and heterologous primary isolates.
Human Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Hu-96T | DL Develop | 96T | 392 EUR |
Mouse Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Mu-48T | DL Develop | 48T | 435 EUR |
Mouse Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Mu-96T | DL Develop | 96T | 561 EUR |
Porcine Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-p-48T | DL Develop | 48T | 493 EUR |
Porcine Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-p-96T | DL Develop | 96T | 641 EUR |
Rat Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Ra-48T | DL Develop | 48T | 310 EUR |
Rat Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Ra-96T | DL Develop | 96T | 389 EUR |
Rabbit Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Rb-48T | DL Develop | 48T | 454 EUR |
Rabbit Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Rb-96T | DL Develop | 96T | 587 EUR |
Sheep Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Sh-48T | DL Develop | 48T | 522 EUR |
Sheep Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Sh-96T | DL Develop | 96T | 681 EUR |
Monkey Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Si-48T | DL Develop | 48T | 522 EUR |
Monkey Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Si-96T | DL Develop | 96T | 681 EUR |
Bovine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-b-48Tests | Reddot Biotech | 48 Tests | 516 EUR |
Bovine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-b-96Tests | Reddot Biotech | 96 Tests | 716 EUR |
Canine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-c-48Tests | Reddot Biotech | 48 Tests | 493 EUR |
Canine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-c-96Tests | Reddot Biotech | 96 Tests | 683 EUR |
Chicken Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Ch-48Tests | Reddot Biotech | 48 Tests | 470 EUR |
Chicken Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Ch-96Tests | Reddot Biotech | 96 Tests | 651 EUR |
Equine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Eq-48Tests | Reddot Biotech | 48 Tests | 528 EUR |
Equine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Eq-96Tests | Reddot Biotech | 96 Tests | 733 EUR |
Human Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Hu-48Tests | Reddot Biotech | 48 Tests | 302 EUR |
Human Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Hu-96Tests | Reddot Biotech | 96 Tests | 409 EUR |
Mouse Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Mu-48Tests | Reddot Biotech | 48 Tests | 447 EUR |
Mouse Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Mu-96Tests | Reddot Biotech | 96 Tests | 618 EUR |
Porcine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-p-48Tests | Reddot Biotech | 48 Tests | 516 EUR |
Porcine Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-p-96Tests | Reddot Biotech | 96 Tests | 716 EUR |
Rat Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Ra-48Tests | Reddot Biotech | 48 Tests | 300 EUR |
Rat Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Ra-96Tests | Reddot Biotech | 96 Tests | 407 EUR |
Rabbit Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Rb-48Tests | Reddot Biotech | 48 Tests | 470 EUR |
Rabbit Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Rb-96Tests | Reddot Biotech | 96 Tests | 651 EUR |
Sheep Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Sh-48Tests | Reddot Biotech | 48 Tests | 551 EUR |
Sheep Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Sh-96Tests | Reddot Biotech | 96 Tests | 765 EUR |
Monkey Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Si-48Tests | Reddot Biotech | 48 Tests | 551 EUR |
Monkey Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Si-96Tests | Reddot Biotech | 96 Tests | 765 EUR |
Bovine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-b-48Tests | Reddot Biotech | 48 Tests | 494 EUR |
Bovine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-b-96Tests | Reddot Biotech | 96 Tests | 684 EUR |
Canine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-c-48Tests | Reddot Biotech | 48 Tests | 472 EUR |
Canine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-c-96Tests | Reddot Biotech | 96 Tests | 653 EUR |
Chicken Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Ch-48Tests | Reddot Biotech | 48 Tests | 450 EUR |
Chicken Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Ch-96Tests | Reddot Biotech | 96 Tests | 622 EUR |
Equine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Eq-48Tests | Reddot Biotech | 48 Tests | 505 EUR |
Equine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Eq-96Tests | Reddot Biotech | 96 Tests | 700 EUR |
Human Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Hu-48Tests | Reddot Biotech | 48 Tests | 290 EUR |
Human Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Hu-96Tests | Reddot Biotech | 96 Tests | 392 EUR |
Mouse Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Mu-48Tests | Reddot Biotech | 48 Tests | 429 EUR |
Mouse Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Mu-96Tests | Reddot Biotech | 96 Tests | 591 EUR |
Porcine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-p-48Tests | Reddot Biotech | 48 Tests | 494 EUR |
Porcine Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-p-96Tests | Reddot Biotech | 96 Tests | 684 EUR |
Rat Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Ra-48Tests | Reddot Biotech | 48 Tests | 288 EUR |
Rat Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Ra-96Tests | Reddot Biotech | 96 Tests | 390 EUR |
Rabbit Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Rb-48Tests | Reddot Biotech | 48 Tests | 450 EUR |
Rabbit Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Rb-96Tests | Reddot Biotech | 96 Tests | 622 EUR |
Sheep Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Sh-48Tests | Reddot Biotech | 48 Tests | 527 EUR |
Sheep Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Sh-96Tests | Reddot Biotech | 96 Tests | 731 EUR |
Monkey Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Si-48Tests | Reddot Biotech | 48 Tests | 527 EUR |
Monkey Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Si-96Tests | Reddot Biotech | 96 Tests | 731 EUR |
Guinea pig Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Gu-48T | DL Develop | 48T | 474 EUR |
Guinea pig Interleukin 4 (IL4) ELISA Kit |
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| DLR-IL4-Gu-96T | DL Develop | 96T | 614 EUR |
Guinea pig Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Gu-48Tests | Reddot Biotech | 48 Tests | 493 EUR |
Guinea pig Interleukin 4 (IL4) ELISA Kit |
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| RDR-IL4-Gu-96Tests | Reddot Biotech | 96 Tests | 683 EUR |
Guinea pig Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Gu-48Tests | Reddot Biotech | 48 Tests | 472 EUR |
Guinea pig Interleukin 4 (IL4) ELISA Kit |
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| RD-IL4-Gu-96Tests | Reddot Biotech | 96 Tests | 653 EUR |
GMP-140/ Rat GMP- 140 Elisa Kit |
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| ELA-E0602r | Lifescience Market | 96 Tests | 886 EUR |
Cyclic-di-GMP |
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| HY-107780 | MedChemExpress | 10mM/1mL | 3712 EUR |
Cyclic GMP Antibody |
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| abx022640-02ml | Abbexa | 0.2 ml | 940 EUR |
Cyclic di-GMP |
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| B7839-.5 | ApexBio | 500 µg | 181 EUR |
Cyclic di-GMP |
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| B7839-1 | ApexBio | 1 mg | 302 EUR |
EGF GMP, CF |
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| PR15112CF | Neuromics | 200 ug | 401 EUR |
SCF GMP, CF |
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| PR15113CF | Neuromics | 50 ug | 1406 EUR |
BDNF GMP, CF |
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| PR15134CF | Neuromics | 5 ug | 636 EUR |
Cyclic-di-GMP |
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| B2045-1000 | Biovision | 555 EUR | |
Cyclic-di-GMP |
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| B2045-250 | Biovision | 207 EUR | |
anti-GMP Synthase |
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| YF-PA25201 | Abfrontier | 50 ul | 334 EUR |
anti-GMP Synthase |
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| YF-PA15898 | Abfrontier | 50 ug | 363 EUR |
anti-GMP Synthase |
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| YF-PA15899 | Abfrontier | 100 ug | 403 EUR |
Cyclic GMP EIA Kit |
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| SKT-211-96 | Stressmarq | 1 plate of 96 wells | 474 EUR |
IL-4 GMP, CF |
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| PR15114CF | Neuromics | 50 ug | 1615 EUR |
IL-7 GMP, CF |
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| PR15115CF | Neuromics | 25 ug | 1693 EUR |
IL-3 GMP, CF |
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| PR15117CF | Neuromics | 50 ug | 1341 EUR |
IL-6 GMP, CF |
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| PR15118CF | Neuromics | 50 ug | 1732 EUR |
IL-15 GMP, CF |
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| PR15119CF | Neuromics | 5 ug | 505 EUR |
NT-3 GMP, CF |
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| PR15120CF | Neuromics | 5 ug | 584 EUR |
Wnt-3a GMP, CF |
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| PR15121CF | Neuromics | 10 ug | 649 EUR |
Activin A GMP, CF |
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| PR15122CF | Neuromics | 50 ug | 1654 EUR |
IFN-gamma GMP, CF |
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| PR15124CF | Neuromics | 100 ug | 884 EUR |
TNF-alpha GMP, CF |
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| PR15126CF | Neuromics | 100 ug | 1497 EUR |
Cyclic GMP Standard, 125UL |
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| C080-125UL | Arbor Assays | 125UL | 123 EUR |
Cyclic GMP Standard, 350UL |
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| C080-350UL | Arbor Assays | 350UL | 227 EUR |
Cyclic GMP Standard, 625UL |
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| C080-625UL | Arbor Assays | 625UL | 227 EUR |
Immunoassay involves a large panel of monoclonal antibodies to characterize lymphocyte phenotype, cell proliferation with mitogen and antigens (tetanus toxoid; GMP C. Albicans, p24 and HIV gp160), HIV-specific cytotoxicity (env and gag), and rate of spontaneous cell mortality. Lymphocyte culture supernatants (stimulated with PHA) are collected to measure cytokines (IL2; IL4 ; IL10; gamma IFN) and T cell clones grown to characterize cytokine phenotype and pattern. Here we provide only a summary of the immunological data collected.