Human Milk
Conditions
Keywords
sample collection
Brief summary
This study will develop a protocol for remote human milk collection for use in gene expression research through application of ribonucleic acid (RNA) preservation reagents. The investigators hypothesize that addition of preservation reagents will preserve RNA in human milk, facilitating downstream analyses that require intact RNA. 50 lactating participants will be recruited and requested to provide reagent-mixed samples to be sent to the university through shipping services where they will be evaluated for RNA quantity, quality and profile.
Detailed description
To preserve the quality of RNAs, commonly used methods for collecting fresh human milk samples typically require a stringent collection and handling routine. This includes proximity to a laboratory (within 20 minutes of driving distance) and ultra-cold storage conditions (below -80°C). Compared to "non-research" settings, current standards in sample collection require either 1) participant travel or 2) home visits for biospecimen collection. These requirements impede the inclusion of wide, non- biased milk samples from a range of populations, which is necessary for robust human subject studies. Subsequently, certain populations are excluded due to arbitrary factors like location, daytime availability, and residence. The development of human milk collection protocols that enable wide-ranging participation is a research priority that supports our research at the University of Wisconsin - Madison and any global investigative efforts that include human milk RNA sequencing. Recruitment will take place in 2 phases (N=25 participants each) to allow for adjustments to the biospecimen collection kit and optimization of RNA integrity. During the first recruitment phase, N=25 participants will be recruited and enrolled using a reagent: human milk concentration of approximately 2:1. Total volume of human milk collection will not exceed 30 mL.
Interventions
DEVICERemote Human Milk Sample Collection
Two products will be evaluated for their ability to preserve RNA in human milk samples. * RNAlater (ThermoFisher©) (ratio variable) * NucleoProtect RNA (Takara©) (ratio variable)\* * Product volume mixed with human milk samples will vary by product and may be titrated as part of protocol development.
Sponsors
University of Wisconsin, Madison
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
OTHER
Masking
NONE
Eligibility
Sex/Gender
FEMALE
Age
18 Years to No maximum
Healthy volunteers
Yes
Inclusion criteria
* English speaking and reading * Lactating for greater than or equal to 4 weeks
Exclusion criteria
* not meeting personal lactation goals * resides outside of Wisconsin
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Successful Development of a Remote Collection Protocol reported as the final Reagent:Sample ratio | through study completion, anticipated to be 1 year | The primary endpoint is generation of a remote collection protocol for use with a leading reagent in RNA preservation of human milk. There is no statistical test or null hypothesis associated with this endpoint, rather, a summary of relevant findings from all study secondary and exploratory endpoints. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Concentration of RNA in Human Milk | through study completion, anticipated to be 1 year | The concentration of extracted RNA (ng/uL) will be measured by a Qubit Fluorometer. |
| Purity of RNA in Human Milk | through study completion, anticipated to be 1 year | The purity of extracted RNA will be measured by a Qubit Fluorometer. Purity will be assessed using wavelengths, the ratio of absorbance for A260/280 and A230/280. |
| RNA Integrity Number (RIN) | through study completion, anticipated to be 1 year | Integrity of the extracted RNA will be measured using an agilent bioanalyzer. RIN is reported on a scale of 1 to 10 where higher numbers indicate higher integrity. |
| Quantitative PCR (qPCR) | through study completion, anticipated to be 1 year | The epithelial cell and microRNA population will be quantified by measuring expression of epithelial cell and microRNA target genes. |
| Sample Collection Feasibility Data | data collected at time of sample return from participant (one time point greater than one month after lactation begins and within 7 days of sample collection) | Participants answer 8-items scored on a 1 (discourage) to 10 (encourage) scale regarding participation in lactation research to assess factors that would facilitate their participation (use of own breast pump, ability to use breast pump, opportunity to privately express a sample in a clinic or at home, handling reagents with gloves, opportunity to participate in maternal and infant health research, opportunity to participate in genetic research). |
Countries
United States
Contacts
PRINCIPAL_INVESTIGATORKatelyn Desorcy-Scherer, PhD, RN
UW School of Medicine and Public Health
Outcome results
None listed