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Cell culture device

A cell culture and micropore technology, applied in tissue cell/virus culture devices, biochemical instruments, biochemical equipment and methods, etc.

Active Publication Date: 2017-04-26
STEMCELL TECHNOLOGIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Colony-forming cell (CFC) assays of non-adherent cells are typically performed in a semi-solid or gel-like medium that prevents convective fluid flow from moving the cells, and thus limits the distance that daughter cells can move from the location of the parent cell

Method used

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  • Cell culture device
  • Cell culture device
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] Example 1 - Production of cell culture device

[0110] Cell culture devices as described above were prepared by fabricating microwells as inserts and inserting them into culture dishes. Some micropores contain submicropores and some do not. The microwells are made to have various maximum dimension to depth ratios, as described above.

[0111] Female molds of several microporous configurations were produced by CNC machining of solid aluminum discs. These circular molds were 35 mm in diameter and 20 mm thick and exhibited a reverse topology surface with micropores. Microporous inserts are produced by casting a polydimethylsiloxane (PDMS)-based elastomer in a mold, followed by curing the elastomer to form a flexible disk containing microwells. Specifically, the elastomer was prepared from a 10:1 (w / w) homogeneous mixture of SylGard 184 (Dow Corning) elastomer and curing agent. This mixture of silicone components was exposed to vacuum (<10 mTorr, 1 hour) to remove any v...

Embodiment 2

[0115] Example 2 - Effect of Pore Configuration on the Immobilization of Particles

[0116] Cell culture devices with microwells of different sizes were evaluated for their ability to compartmentalize particles. The cell culture devices tested in this example did not contain submicrowells. Evaluation was accomplished using fluorescent polystyrene microparticles (Bangs FS06F, 7.3 μm diameter). The suspension of microparticles is placed in several individual wells of several cell culture devices. All wells are square at the top of the well. The pores have the following dimensions:

[0117] structure Wall angle (degrees) Pore ​​Width (mm) Microhole Depth(mm) Maximum size to depth ratio A 15 1.0 1.0 1.4∶1 B 15 0.8 0.5 2.3∶1 C 37 0.8 0.38 3.0∶1

[0118] The particles were allowed to settle by gravity into the wells; the remaining wells were left empty containing only phosphate buffered saline (PBS). The cell culture device is then ...

Embodiment 3

[0121] Example 3 - Effect of Well Configuration on Accommodating Cell Colonies

[0122] Human hematopoietic progenitor cells in liquid medium were seeded into cell culture dishes containing microwells of various configurations. The cell culture devices tested in this example did not contain submicrowells. All wells are square at the top of the well. The dimensions of the microwells are summarized in the table below:

[0123] structure Wall angle (degrees) Pore ​​Width (mm) Microhole Depth(mm) Maximum size to depth ratio A 37 0.8 0.38 3.0∶1 B 30 1 0.75 1.9∶1 C 20 1 0.75 1.9∶1 D 15 1 0.75 1.9∶1 E 15 1 1 1.4∶1

[0124] With about 7 colonies / cm 2 The cell culture device was seeded at a colony density of , and the cells were sedimented into the microwells by slow centrifugation. Seed the cell culture device subsequently in a permissive culture environment at 37 °C and containing 5% CO 2 incubated in a humidified atm...

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Abstract

A cell culture device comprises holes. The holes are provided with many micro holes and the first public fluid volume is above the micro holes. Each micro hole includes a group of sub-micro holes and the second public fluid volume is above the group of sub-micro hole in each micro hole.

Description

technical field [0001] The present disclosure relates to cell culture devices. In particular, the present disclosure relates to cell culture devices, such as multiwell plates, that can be used in colony formation assays. Background technique [0002] What follows is not an admission that anything disclosed below is prior art or part of the common general knowledge of those skilled in the art. [0003] Colony-forming cell (CFC) assays of non-adherent cells are typically performed in a semi-solid or gel-like medium that prevents convective fluid flow from moving the cells, and thus limits the distance that daughter cells move from the location of the parent cell. This results in the formation of multicellular colonies derived from a single cell as the daughter cells continue to divide. Colony formation assays can provide quantitative information on the number of individual viable progenitor cells in a sample and allow isolation and sampling of individual colonies for subclon...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00
CPCC12M25/04
Inventor O·艾格勒S·伍德赛德
Owner STEMCELL TECHNOLOGIES