Primary Cell Harvesting
Primary Cell Harvesting - Related Videos
06:44
Harvest and Primary Culture of Leptomeningeal Lymphatic Endothelial Cells
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Cited by 2 •
2023
Leptomeningeal lymphatic endothelial cells (LLECs), a recently identified intracranial cell type, have poorly understood functions. This study presents a reproducible protocol for harvesting LLECs from mice and establishing in vitro primary cultures. This protocol is designed to enable researchers to delve into the cellular functions and potential clinical implications of LLECs.
07:06
Primary Clarification of CHO Harvested Cell Culture Fluid using an Acoustic Separator
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Cited by 7 •
2020
Presented here is a protocol for the primary clarification of CHO cell culture using an acoustic separator. This protocol can be used for the primary clarification of shake flask cultures or bioreactor harvests and has the potential application for continuous clarification of the cell bleed material during perfusion bioreactor operations.
04:09
Isolating RPE and IPE Cells from Bovine Eye: A Procedure to Harvest and Culture Bovine Primary Pigment Epithelial Cells
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2025
This video demonstrates the procedure to isolate the retinal pigment epithelial, or RPE, cells and iris pigment epithelial, or IPE, cells from harvested bovine eyes. The isolated cells can be cultured to study their role in the treatment of ocular diseases.
02:06
Harvesting Adeno-Associated Virus from Transfected Mammalian Cells
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2026
Source: Rghei, A. D., et al. Production of Adeno-Associated Virus Vectors in Cell Stacks for Preclinical Studies in Large Animal Models. J. Vis. Exp. (2021)This video demonstrates the recovery of crude adeno-associated virus from transfected mammalian cells using chemical lysis and centrifugation, enabling high-yield AAV production for downstream purification and preclinical gene therapy applications.
10:02
piggyBac Transposon System Modification of Primary Human T Cells
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Cited by 16 •
2012
We describe a method to genetically modify primary human T cells with a transgene using the non-viral piggyBac transposon system. T cells modified to using the piggyBac transposon system exhibit stable transgene expression.
FAQs
Viability depends on coordinating mechanical disaggregation, enzymatic digestion, filtration, and centrifugation rather than relying on one step alone. Mechanical treatment breaks tissue into smaller fragments, while enzymes help release cells from the extracellular matrix. Subsequent separation steps prepare the population for culture, where suitable selective conditions can support survival and enrichment of the desired cells.
Enzymatic digestion helps loosen the extracellular matrix that holds cells within tissue. This complements mechanical disaggregation by improving cell release without treating tissue breakdown as purely physical. The balance matters because the harvesting workflow must expose individual cells for downstream use while maintaining sufficient viability to support culture and engineering applications.
Yield and purity are shaped by the effectiveness of tissue disaggregation, matrix digestion, filtration, centrifugation, and selective culture conditions. These steps determine how many cells are recovered and how strongly the desired population is enriched. Donor variation also contributes to differences between preparations, making consistency a central challenge when primary cells enter engineered systems.
Primary cells retain many properties of their source tissue, which can improve the physiological relevance of tissue-engineered constructs, organoids, and disease models. Their value comes with practical constraints: donor-to-donor variation, variable yield and purity, and limited expansion capacity can complicate efforts to produce uniform or large quantities of material.
A typical workflow begins with mechanical disaggregation of tissue and enzymatic digestion of its extracellular matrix. The resulting mixture then undergoes filtration and centrifugation before cells are placed under selective culture conditions. Together, these stages support physical preparation, separation, viability preservation, and enrichment of the population needed for subsequent laboratory or engineering work.
Selective culture conditions help enrich the desired cell population after tissue has been mechanically and enzymatically processed. This is important because harvesting produces material that may require further selection before use. Enrichment can make the resulting cells more suitable for controlled engineering studies, although it does not remove challenges associated with donor variation or limited expansion.
Engineers use harvested primary cells as biologically relevant starting material for tissue-engineered constructs, regenerative medicine studies, organoid development, and disease models. Because the cells preserve characteristics associated with source tissue, they can strengthen the physiological relevance of these systems. Their limited expansion capacity, however, must be considered when planning experiments or construct production.