Micro-fluidic device for selective sorting of highly motile and morphologically normal sperm from unprocessed semen

Patent metadata

Field	Value
Publication number	US-11618882-B2
Application number	US-201716070368-A
Country	US
Kind code	B2
Filing date	Jan 22, 2017
Priority date	Jan 22, 2016
Publication date	Apr 4, 2023
Grant date	Apr 4, 2023

Abstract

A microfluidic chip is provided for self-sorting highly motile, morphologically normal sperm cell with high DNA integrity from a fresh semen sample. The sperm self-sorting microfluidic chip has one or more inlet chambers, and sperm collection outlet chamber(s), and the middle of the channel features various micro-fabricated structures in different geometrical shapes and orientations, with varying periodicities and patterns, such as an array of micro-fabricated pillars that facilitate the transport of the active and healthy sperm into the outlet chamber.

First claim

What is claimed is:

1. A method of self-sorting motile, morphologically-normal sperm cells with high DNA integrity from raw or unprocessed semen, comprising:
a) providing a fluidic channel with an inlet at one end and an outlet at the other end,

wherein an array of pillar-structures is periodically spaced inside the fluidic channel, wherein the spacing between adjacent pillar-structures ranges from one micrometer to 250 micrometers;
b) introducing raw or unprocessed semen containing sperm cells at the inlet of the fluidic channel;
c) self-sorting the sperm cells by their own self-induced movements within the fluidic channel through their interactions with the periodically spaced array of pillar-structures and without the use of any external flow, forces or mechanisms to feed the raw or unprocessed sperm through the fluidic channel; and
d) collecting at the outlet sorted sperm cells from the fluidic channel, and

wherein the sorted sperm cells are the motile, morphologically-normal sperm cells with high DNA integrity compared to the raw or unprocessed semen.

2. The method of claim 1 , wherein a length of the fluidic channel ranges from 4 mm to 20 mm, and wherein a height of the pillar-structures ranges from 20 μm to 80 μm.

3. The method of claim 1 , wherein the spacing between adjacent pillar-structures ranges from 5 μm to 200 μm.

4. The method of claim 1 , wherein the spacing between adjacent pillar-structures ranges from 5 μm to 30 μm.

5. The method of claim 1 , wherein the spacing between adjacent pillar-structures ranges from 18 μm to 30 μm in a first direction.

6. The method of claim 1 , wherein the length of the fluidic channel ranges from 12 mm to 20 mm.

7. The method of claim 1 , wherein the width of the fluidic channel is 1.5 mm.

8. The method of claim 1 , wherein allowing the sperm cells in the sample to move from the inlet, through the array and to the outlet comprises incubating for a period of time ranging from 5 minutes to 30 minutes.

9. The method of claim 1 , wherein the array of pillar-structures has a periodicity of 18×26 μm, 22×22 μm, 22×26 μm, 26×26 μm or 30×26 μm.

10. The method of claim 5 , wherein the spacing between adjacent pillar-structures ranges from 22 μm to 26 μm in a second direction.

Assignees

• Univ Leland Stanford Junior
• Worcester Polytech Inst

Inventors

• Demirci Utkan
• Tüzel Erkan
• Kingsley James Leonard
• Chinnasamy Thiruppathiraja

Classifications

• C12N5/0612 Primary

  sorting of gametes, e.g. according to sex or motility · CPC title

• C12N5/061

  Sperm cells, spermatogonia · CPC title

• C12M47/04

  Cell isolation or sorting (purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04) · CPC title

• C12M23/16

  Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093) · CPC title

• B01L2300/0832

  cylindrical, tube shaped · CPC title