Periodic Reporting for period 1 - MicroFSMA (MicroFluidic based platform for SeMen Analysis)
Okres sprawozdawczy: 2019-07-01 do 2021-06-30
MicroFSMA aimed to facilitate a microfluidic-based device for quality sperm separation. Additionally, the project targeted to facilitate the user-friendly computational framework for automated semen analysis.
Human infertility is a growing health concern. The assisted reproductive techniques (ARTs), which involve “Intra Cytoplasmic Sperm Injection (ICSI)”, “Intrauterine Insemination (IUI)”, and “In-vitro Fertilization (IVF)” are being practiced in treating male/female infertility. It has been two decades of research; the success rate of ARTs is saturated to merely ~35% approximately. Many previous studies claim that sperm selection is one of the critical steps for ARTs. The conventional methods including swim-up and density centrifugation cause DNA fragmentation and that leads to one of the potential causes for the ART’s failure.
The market for ARTs services, devices, and related consumable is growing at a promising compound annual growth rate (CAGR). Modification in human reproduction laws and increased infertility percentage are the potential market drivers. On the contrary, the higher cost and failure rates appeared as potential market threats. Hence, technological advancement and rapid commercialization is an urgent call from the subject research theme.
Previous studies claim that the sperm selection techniques inspired by natural swimming mechanisms viz rheotaxis, thermotaxis, chemotaxis can improve the success rate of ARTs.
There are two main technical objectives of the MicroFSMA: 1) Developed a microfluidic chip that is capable in mimicking the physiology and micro-environment of the female reproductive tract while hosting the semen deposition. 2). Developed an automated computational framework that enables the rapid assessment of the parameters associated with sperm cell kinematics (curvilinear velocity (VCL), average path velocity, straight line velocity (VSL), linearity (LIN), wobbling (WOB), and beat frequency (BCF)).
Human infertility is a growing health concern. The assisted reproductive techniques (ARTs), which involve “Intra Cytoplasmic Sperm Injection (ICSI)”, “Intrauterine Insemination (IUI)”, and “In-vitro Fertilization (IVF)” are being practiced in treating male/female infertility. It has been two decades of research; the success rate of ARTs is saturated to merely ~35% approximately. Many previous studies claim that sperm selection is one of the critical steps for ARTs. The conventional methods including swim-up and density centrifugation cause DNA fragmentation and that leads to one of the potential causes for the ART’s failure.
The market for ARTs services, devices, and related consumable is growing at a promising compound annual growth rate (CAGR). Modification in human reproduction laws and increased infertility percentage are the potential market drivers. On the contrary, the higher cost and failure rates appeared as potential market threats. Hence, technological advancement and rapid commercialization is an urgent call from the subject research theme.
Previous studies claim that the sperm selection techniques inspired by natural swimming mechanisms viz rheotaxis, thermotaxis, chemotaxis can improve the success rate of ARTs.
There are two main technical objectives of the MicroFSMA: 1) Developed a microfluidic chip that is capable in mimicking the physiology and micro-environment of the female reproductive tract while hosting the semen deposition. 2). Developed an automated computational framework that enables the rapid assessment of the parameters associated with sperm cell kinematics (curvilinear velocity (VCL), average path velocity, straight line velocity (VSL), linearity (LIN), wobbling (WOB), and beat frequency (BCF)).
The project was commenced with reviewing the niche for such a technology. We have conducted a TwitterTM sentiment analysis to assess the frustration within the ARTs customer. Additionally, our interaction with the end-user (reproductive clinical practitioner) hinted us to develop a user-friendly, inexpensive, rapid, and centrifugation-free technology for quality sperm isolation. The literature and patent survey were conducted and reasons behind the slow commercialization rate for microfluidic-based technology were extracted. Lack of industry-academia interaction, material choices for prototyping, and the cost of regulatory approvals are the chief hurdles in the commercialization pathway.
An independent webpage known as “Phlagella” was created to understand the market size, customer segment, cost structure. The “Phlagella” web portal comprises four web pages where project description, ER bio-sketch, contact, and knowledge associated with artificial reproductive techniques (ARTs) and sperm navigation were shared to the layman audience. The social media platforms (Medium©, Twitter™, Facebook©), was exploited for the blogs dissemination. and written blogs’ articles were propagated through the created social profiles. Apart from the project description, five blog articles circulated for the layman audiences:
• Spermatozoa: one of the reasons for our existence
• Artificial Reproductive Techniques (ARTs): a ray of hope in treating infertility.
• Practices in sperm selection for artificial reproductive techniques: Are we doing right?
• Mechanism of sperm swimming in the female reproductive tract: what can we learn from mother nature?
• Emerging technologies in sperm selection: Can we solve ARTs frustration?
Assessing the technological void and demand from the end-user, a microfluidic device and biological protocol for quality sperm separation were developed. The chip replicates the physiology and microenvironment of the female reproductive tract and enables the centrifugation-free separation of quality sperm cells from the semen specimen. The chip convolves the passive fluid-flow methodology and comprises user-friendly biological protocols for high-throughput quality sperm cell collection. The chip characterization for the flow. The experiment was conducted with the polystyrene microparticles, and a time interval was identified for the cell separation protocols (approximately 30 minutes and upstream flow interval varies [150-20] µm/s for the mentioned duration). The temperature gradient along the fluidic channel was generated via pasting the Kapton tape. The temperature control of the Kapton tape was achieved through the coupling of JT-sensors, AD cards, and microcontrollers.
We have also developed an automated tracking tool for rapid sperm kinematic parameter extraction. The tool involves image processing of the sperm swimming where image denoising, segmentation, and object detection were executed. Further, a multi-object tracking algorithm was implemented to extract the trajectories of swimming sperm heads. Obtained trajectories from the abovementioned modules were saved in .XLSX extension. Further, the prepared programming function was executed to evaluate “Computer Assisted Semen Analysis (CASA)”. The CASA parameters including curvilinear velocity (VCL), average path velocity (VAP), progressive velocity (VSL), linearity (LIN), wobbling (WOB), progressiveness (PROG), and beat frequency (BCF). Developed module estimates mentioned motility parameters. The validation was conducted against the ImageJ CASA plugin. However, the validation was not straightforward as the CASA plugin did not provide sufficient accuracy over the input images. Hence, we conducted the ImageJ CASA plugin study with our post-processed image data. The coherency in results was obtained.
The dissemination and published results of the above-mentioned technical outcomes will be updated on the Béez Biotech webpage.
Apart from technical advancement, the positive development of MicroFSMA and the acquisition of entrepreneurial knowledge have encouraged ER to create Béez Biotech SAS. Cherry has been supportive and encouraging through the incorporation of the enterprise. The incorporation of Béez Biotech has been accomplished and SIREN and Kbis (French business incorporation certificate) and the company will be active right after the end of the MicroFSMA grant. The mission of the Béez Biotech is to translate the no-technical and technical achievements of MicroFSMA and deliver a user-friendly and inexpensive commercial solution for quality sperm cell selection.
An independent webpage known as “Phlagella” was created to understand the market size, customer segment, cost structure. The “Phlagella” web portal comprises four web pages where project description, ER bio-sketch, contact, and knowledge associated with artificial reproductive techniques (ARTs) and sperm navigation were shared to the layman audience. The social media platforms (Medium©, Twitter™, Facebook©), was exploited for the blogs dissemination. and written blogs’ articles were propagated through the created social profiles. Apart from the project description, five blog articles circulated for the layman audiences:
• Spermatozoa: one of the reasons for our existence
• Artificial Reproductive Techniques (ARTs): a ray of hope in treating infertility.
• Practices in sperm selection for artificial reproductive techniques: Are we doing right?
• Mechanism of sperm swimming in the female reproductive tract: what can we learn from mother nature?
• Emerging technologies in sperm selection: Can we solve ARTs frustration?
Assessing the technological void and demand from the end-user, a microfluidic device and biological protocol for quality sperm separation were developed. The chip replicates the physiology and microenvironment of the female reproductive tract and enables the centrifugation-free separation of quality sperm cells from the semen specimen. The chip convolves the passive fluid-flow methodology and comprises user-friendly biological protocols for high-throughput quality sperm cell collection. The chip characterization for the flow. The experiment was conducted with the polystyrene microparticles, and a time interval was identified for the cell separation protocols (approximately 30 minutes and upstream flow interval varies [150-20] µm/s for the mentioned duration). The temperature gradient along the fluidic channel was generated via pasting the Kapton tape. The temperature control of the Kapton tape was achieved through the coupling of JT-sensors, AD cards, and microcontrollers.
We have also developed an automated tracking tool for rapid sperm kinematic parameter extraction. The tool involves image processing of the sperm swimming where image denoising, segmentation, and object detection were executed. Further, a multi-object tracking algorithm was implemented to extract the trajectories of swimming sperm heads. Obtained trajectories from the abovementioned modules were saved in .XLSX extension. Further, the prepared programming function was executed to evaluate “Computer Assisted Semen Analysis (CASA)”. The CASA parameters including curvilinear velocity (VCL), average path velocity (VAP), progressive velocity (VSL), linearity (LIN), wobbling (WOB), progressiveness (PROG), and beat frequency (BCF). Developed module estimates mentioned motility parameters. The validation was conducted against the ImageJ CASA plugin. However, the validation was not straightforward as the CASA plugin did not provide sufficient accuracy over the input images. Hence, we conducted the ImageJ CASA plugin study with our post-processed image data. The coherency in results was obtained.
The dissemination and published results of the above-mentioned technical outcomes will be updated on the Béez Biotech webpage.
Apart from technical advancement, the positive development of MicroFSMA and the acquisition of entrepreneurial knowledge have encouraged ER to create Béez Biotech SAS. Cherry has been supportive and encouraging through the incorporation of the enterprise. The incorporation of Béez Biotech has been accomplished and SIREN and Kbis (French business incorporation certificate) and the company will be active right after the end of the MicroFSMA grant. The mission of the Béez Biotech is to translate the no-technical and technical achievements of MicroFSMA and deliver a user-friendly and inexpensive commercial solution for quality sperm cell selection.
From ER perspective, MicroFSMA is one of the first technology that replicates the swimming mechanism of the spermatozoa in a microfluidic chip. As currently, Béez Biotech is looking forward to patenting generated intellectual property.
The positive social outcomes of MicroFSMA technology are still far away. The Béez Biotech has taken the responsibility to carry forward the project and tackle the reproductive clinician's demands to upsurge the ARTs consumer expectations.
The positive social outcomes of MicroFSMA technology are still far away. The Béez Biotech has taken the responsibility to carry forward the project and tackle the reproductive clinician's demands to upsurge the ARTs consumer expectations.