Immunosurgery Protocol
Immunosurgery Protocol - Related Videos
11:56
Derivation of Human Embryonic Stem Cells by Immunosurgery
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Cited by 12 •
2007
The ability of human embryonic stem cells to self-renew and differentiate into all cell types of the body suggests that they hold great promise for both medical applications and as a research tool for addressing fundamental questions in development and disease. Here, we provide a concise, step-by-step protocol for the derivation of human embryonic stem cells from embryos by immunosurgical isolation of the inner cell mass.
07:12
Neutrophil Isolation Protocol
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Cited by 173 •
2008
Neutrophils are among the first cells to arrive on the site of inflammatory immune response, and their functions and mechanisms have been studied extensively in vitro. We demonstrate a standard density gradient separation method to isolate human neutrophils from whole blood using commercially available separation media.
07:56
Parabiosis in Mice: A Detailed Protocol
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Cited by 175 •
2013
Parabiotic joining of two organisms leads to the development of a shared circulatory system. In this protocol, we describe the surgical steps to form a parabiotic connection between a wild-type mouse and a constitutive GFP-expressing mouse.
07:24
A Protocol for the Production of KLRG1 Tetramer
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Cited by 2 •
2010
This protocol describes the production of KLRG1 tetramer, which is a powerful tool for the analysis of KLRG1 ligands.
13:03
Protocol for Mosquito Rearing (A. gambiae)
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Cited by 64 •
2007
This video illustrates the general techniques used to rear Anopheles gambiae in the laboratory. The methods for caring for laboratory mosquitoes are demonstrated through all stages of the organism's life cycle from larvae to pupae to blood-feeding adults.
FAQs
Selectivity comes from differential antibody recognition. Antibodies bind surface antigens on the troph ectoderm, marking those outer cells for immune-mediated injury, while the inner cell mass remains relatively protected. This molecular targeting allows the two blastocyst compartments to be separated according to their surface properties rather than by nonspecific destruction of the entire embryo.
Complement acts as the destructive effector in the protocol. Once antibodies are attached to antigens on trophoectoderm cells, complement activation damages their membranes and leads to cell lysis. This step converts antibody binding into physical removal of the surrounding layer, making recovery of the protected inner cell mass possible.
The inner cell mass is preserved because the immune targeting is directed primarily toward surface antigens on the surrounding trophoectoderm. With less effective antibody-mediated complement injury, its cells remain available for recovery after the outer layer lyses. That relative protection is essential for obtaining material suitable for subsequent culture and developmental analysis.
Surface-antigen distribution is the key distinguishing feature. Cells displaying the targeted antigens can bind the antibodies and become vulnerable to complement-mediated membrane damage, whereas cells lacking comparable exposure are less affected. In this way, the protocol links cellular identity to differential immune susceptibility, enabling selective enrichment of the desired compartment.
The workflow first exposes the blastocyst to antibodies that recognize surface antigens on the outer cells. Complement is then activated to damage and lyse that layer. After lysis, the disrupted trophoectoderm is removed, and the remaining inner cell mass is recovered for culture or further developmental study.
The recovered inner cell mass provides material for embryonic stem-cell derivation and for investigating early developmental events. Researchers can examine lineage specification and cell differentiation after the surrounding trophoectoderm has been removed. Consequently, the method connects physical cell isolation with studies of how early mammalian cell populations acquire distinct developmental fates.