Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, the...Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, then incubated them and collected cell supernatant. By using Caprylic acid- saturated Ammonium sulfate method, monoclonal antibody was purified, and IgG subtype identification was conducted by their subtype classification kit of sigma. In traditional way, the preparation of monoclonal antibody cannot do without BABL/C mice and SP2/0 hybridoma cells. Consider this kinds of monoclonal antibody as a positive control, the reactivity and specificity of monoclonal antibodies were identified by Dot-ELISA. Results: By ELISA, we obtained four strains of positive umbilical cord blood cells. After subculture, cryopreservation and resuscitation in vitro, three of them were confirmed to secrete monoclonal antibody against rubella virus stably. The result of monoclonal antibody subtype classification showed that, both IgG1 and IgG2 types were detected. However, the quantity of monoclonal antibody prepared by umbilical cord blood cells was less than that from traditional method. Conclusions: The method of prepare monoclonal antibody using umbilical cord blood cells is feasible.展开更多
Mechanical allodynia(MA),including punctate and dynamic forms,is a common and debilitating symptom suffered by millions of chronic pain patients.Some peripheral injuries result in the development of bilateral MA,while...Mechanical allodynia(MA),including punctate and dynamic forms,is a common and debilitating symptom suffered by millions of chronic pain patients.Some peripheral injuries result in the development of bilateral MA,while most injuries usually led to unilateral MA.To date,the control of such laterality remains poorly understood.Here,to study the role of microglia in the control of MA laterality,we used genetic strategies to deplete microglia and tested both dynamic and punctate forms of MA in mice.Surprisingly,the depletion of central microglia did not prevent the induction of bilateral dynamic and punctate MA.Moreover,in dorsal root ganglion-dorsal root-sagittal spinal cord slice preparations we recorded the low-threshold Aβ-fiber stimulation-evoked inputs and outputs of superficial dorsal horn neurons.Consistent with behavioral results,microglial depletion did not prevent the opening of bilateral gates for Aβpathways in the superficial dorsal horn.This study challenges the role of microglia in the control of MA laterality in mice.Future studies are needed to further understand whether the role of microglia in the control of MA laterality is etiology-or species-specific.展开更多
The original version of this article unfortunately contained one error.The authors found that in the middle panel of Fig.2A,the example traces recorded from a contralateral dorsal horn neuron of the capsaicin-treated ...The original version of this article unfortunately contained one error.The authors found that in the middle panel of Fig.2A,the example traces recorded from a contralateral dorsal horn neuron of the capsaicin-treated mouse were wrong,and the corrected Fig.2 was as follows.展开更多
文摘Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, then incubated them and collected cell supernatant. By using Caprylic acid- saturated Ammonium sulfate method, monoclonal antibody was purified, and IgG subtype identification was conducted by their subtype classification kit of sigma. In traditional way, the preparation of monoclonal antibody cannot do without BABL/C mice and SP2/0 hybridoma cells. Consider this kinds of monoclonal antibody as a positive control, the reactivity and specificity of monoclonal antibodies were identified by Dot-ELISA. Results: By ELISA, we obtained four strains of positive umbilical cord blood cells. After subculture, cryopreservation and resuscitation in vitro, three of them were confirmed to secrete monoclonal antibody against rubella virus stably. The result of monoclonal antibody subtype classification showed that, both IgG1 and IgG2 types were detected. However, the quantity of monoclonal antibody prepared by umbilical cord blood cells was less than that from traditional method. Conclusions: The method of prepare monoclonal antibody using umbilical cord blood cells is feasible.
基金supported by grants from the Ministry of Science and Technology of China(2021ZD0203302)the National Natural Science Foundation of China(32170996,32060199)+3 种基金the Shenzhen-Hong Kong Institute of Brain Science-Shenzhen Fundamental Research Institutions(2021SHIBS0002)the Guangdong Science and Technology Committee(2019A1515010041,A2021319)the Shenzhen Innovation Committee of Science and Technology(ZDSYS20200811144002008)the Shenzhen Science and Technology Innovation Committee(JCYJ20180302174233348).
文摘Mechanical allodynia(MA),including punctate and dynamic forms,is a common and debilitating symptom suffered by millions of chronic pain patients.Some peripheral injuries result in the development of bilateral MA,while most injuries usually led to unilateral MA.To date,the control of such laterality remains poorly understood.Here,to study the role of microglia in the control of MA laterality,we used genetic strategies to deplete microglia and tested both dynamic and punctate forms of MA in mice.Surprisingly,the depletion of central microglia did not prevent the induction of bilateral dynamic and punctate MA.Moreover,in dorsal root ganglion-dorsal root-sagittal spinal cord slice preparations we recorded the low-threshold Aβ-fiber stimulation-evoked inputs and outputs of superficial dorsal horn neurons.Consistent with behavioral results,microglial depletion did not prevent the opening of bilateral gates for Aβpathways in the superficial dorsal horn.This study challenges the role of microglia in the control of MA laterality in mice.Future studies are needed to further understand whether the role of microglia in the control of MA laterality is etiology-or species-specific.
文摘The original version of this article unfortunately contained one error.The authors found that in the middle panel of Fig.2A,the example traces recorded from a contralateral dorsal horn neuron of the capsaicin-treated mouse were wrong,and the corrected Fig.2 was as follows.