Identifying status of male fertility by determining sperm capacitation
This disclosure provides a method for determining male fertility status. The method comprises determining G M1 distribution patterns following induced sperm capacitation, identifying the frequency of distribution of various patterns, and determining if the frequency distribution of certain G M1 patterns in response to induced capacitation is altered. Based on the change in the frequency distribution patterns of certain patterns in response to induced capacitation, alone or in combination with other sperm attributes, male fertility status can be identified.
1. A method for identifying fertility status of a human male individual comprising:
obtaining a sperm sample from a human male;
exposing said sperm to capacitating conditions in vitro to obtain capacitated sperm;
fixing the in vitro capacitated sperm with a fixative;
staining the in vitro capacitated and fixed sperm;
identifying a capacitated state of the human male based on a sum percentage of apical acrosome (AA) G M1 patterns;
and a percentage of acrosomal plasma membrane (APM) G M1 patterns in the in vitro capacitated, fixed, and stained sperm; and
determining a fertility status of the human male by comparing the identified capacitated state to a predetermined fertility threshold based on a sum of a percentage of AA and a percentage of APM G M1 patterns for a fertile population; and
proceeding with a suitable assisted reproduction treatment to achieve fertilization based on the fertility status so determined, wherein the suitable assisted reproduction treatment comprises performing an intrauterine insemination (IUD), an in vitro fertilization (IVF) or an intra-cytoplasmic sperm injection (ICSI).
2. The method of claim 1 , wherein the fixative comprises paraformaldehyde, glutaraldehyde or combinations thereof.
3. The method of claim 1 , wherein the predetermined fertility threshold is the G M1 distribution pattern percentage at which the fertility of a population ceases to substantially increase, and wherein a frequency less than the fertility threshold is indicative of infertile or sub-fertile fertility status.
4. The method of claim 1 , wherein the predetermined fertility threshold is fertile fertility status.
5. The method of claim 1 , wherein the predetermined fertility threshold is subfertile fertility status.
6. The method of claim 1 , wherein the predetermined fertility threshold is infertile fertility status.
7. The method of claim 1 , wherein the predetermined fertility threshold is a value obtained from a dataset comprising a plurality of individuals.
8. The method of claim 1 , wherein the capacitating conditions to which the capacitated sperm have been exposed comprise:
i) centrifugation of a sample comprising the sperm through a density gradient;
ii) suspending a fraction of the density gradient comprising the sperm in a liquid comprising modified human tubal fluid (mHTF);
iii) centrifugation of the fraction of ii) to obtain a pellet comprising the sperm;
iv) suspending the pellet of iii) in a liquid comprising mHTF; and
v) capacitating the sperm of iv) using 2-hydroxy-propyl-β-cyclodextrin.
9. A method for identifying fertility status of a human male individual comprising:
exposing a sperm sample from the individual to capacitating conditions in vitro to obtain capacitated sperm;
fixing the in vitro capacitated sperm with a fixative;
staining the in vitro capacitated and fixed sperm;
identifying G M1 patterns, including induced apical acrosome (AA) G M1 patterns and acrosomal plasma membrane (APM) G M1 patterns;
comparing a frequency of AA+(APM)G M1 patterns in the in vitro capacitated, fixed, and stained sperm to a fertility threshold which is one standard deviation below a mean percentage of AA+APM G M1 patterns for a fertile population,
determining a fertile status of the human male when the frequency of AA+APM G M1 patterns is greater than the fertility threshold, and determining an infertile or a sub-fertile status of the human male when the frequency of AA+APM G M1 patterns is less than the fertility threshold; and
proceeding with a suitable assisted reproduction treatment to achieve fertilization based on the infertile or sub-fertile status so determined, and wherein the suitable assisted reproduction treatment comprises an intrauterine insemination (IUI), an in vitro fertilization (IVF) or an intra-cytoplasmic sperm injection (ICSI).
10. The method of claim 9 , further comprising:
comparing the frequency of AA+APM G M1 patterns in the capacitated sperm to an infertility threshold which is two standard deviations below the mean percentage of AA+APM G M1 patterns for a fertile population; and
determining a fertile status of the human male when the frequency of AA+APM G M1 patterns is greater than the fertility threshold, and determining an infertile or a sub-fertile status of the human male when the frequency of AA+APM G M1 patterns is less than the fertility threshold.
11. The method of claim 9 , wherein exposing the sperm sample from the individual to capacitating conditions in vitro to obtain capacitated sperm comprises:
i) centrifugation of a sample comprising the sperm through a density gradient;
ii) suspending a fraction of the density gradient comprising the sperm in a liquid comprising modified human tubal fluid (mHTF);
iii) centrifugation of the fraction of ii) to obtain a pellet comprising the sperm;
iv) suspending the pellet of iii) in a liquid comprising mHTF; and
v) capacitating the sperm of iv) using hydroxy-propyl-β-cyclodextrin.