Primary Cell Culture of Hepatic Tumorous Cells From Routine Fine-needle Aspiration: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for Primary Cell Culture of Hepatic Tumorous Cells From Routine Fine-needle Aspiration (NCT01549275), a registered phase not stated study. The primary outcome is Correlation Between the Growth Speeds of the Cultured Cells and the AJCC TNM Stage (7th Eds) at Entering of the Study..

2. Clinical Question

The study’s stated purpose was to evaluate the success rate of culturing hepatocellular carcinoma cells and cancer-associated fibroblasts from residual specimens, and to consider the method as a possible additional tool for personalized treatment research. The primary endpoint listed in the registry is narrower: it concerns the association between cultured-cell growth speed and stage at entry.

Rapidly proliferative cultures were defined using either of two criteria at day 28: growth area exceeding two times the area measured at day 14, or growth area reaching > 70% of the flask’s growth area. Special staining was used to distinguish rapid growth of HCC cells alone, HCC cells with concomitant cancer-associated fibroblasts, and fibroblasts alone. The registry record does not report the number of patients meeting these definitions or the comparison results by stage group.

3. Trial Design

The registry describes a prospective study of primary cell culture using residual specimens from routine fine-needle aspiration of hepatic tumors. The listed condition is hepatocellular carcinoma, and the reported enrollment is 105. The trial is marked completed; its recorded start is 2010-04 and its primary completion is 2013-01. The sponsor is Kaohsiung Medical University Chung-Ho Memorial Hospital, and the official title field names Kaohsiung Medical University Hospital.

The primary endpoint compares the incidence of rapidly proliferative cultured cells between two stage categories: AJCC TNM stage IIIA or lower and stage IIIB or higher. The stage classification is identified as the seventh edition. Cultures were evaluated at 28 days after plating, with growth speed defined by growth-area criteria that use measurements at 14 and 28 days. Special staining further categorizes which cell types proliferated rapidly.

This is not a randomized treatment comparison. The registry does not state an allocation method, and the stage categories are observational groups rather than assigned interventions. Consequently, a difference between them would describe an association in the enrolled study population; it would not by itself establish that stage caused a difference in culture growth.

A secondary outcome concerns the relationship between cultured-cell growth speeds and worsening of AJCC TNM stage or HCC-related death 6 months after plating. The registry lists two outcome measures and nine analyses, but does not report the analysis details or numerical findings for these outcomes. The sample-size justification for the original study, including any planned assumptions, is not reported in the registry record.

4. Participant Flow and Accounting

The registry reports how participants moved through each stage of the study. All counts below are exactly as posted; missing counts are not inferred from other stages.

Before assignment: A total of 114 patients signed the inform consents. However, 9 patients were excluded due to negative cytological and pathologic results of malignancy in 4 patients, adenocarcinoma in 2 patients and fungal contamination during the preparation of the specimens in 3 patients.

Overall Study

Reasons for not completing this period:

Analysis population for the primary outcome, as defined in the registry: Comparison the incidence of patients with rapidly proliferative cultured cells between two groups

5. Baseline Characteristics

The table reproduces the baseline characteristics posted in the registry, which describe the participants analysed at baseline and help show whether the randomized groups were comparable.

6. Primary Endpoint

The registry lists the primary outcome as Correlation Between the Growth Speeds of the Cultured Cells and the AJCC TNM Stage (7th Eds) at Entering of the Study., measured over 28 days after plating of cells.

The primary endpoint concerns whether rapidly proliferative cultured cells were more or less common across the two AJCC TNM stage groups at study entry. It is a binary participant-level outcome: each patient is classified according to whether the cultured cells meet the registry’s rapid-proliferation definition. The record describes two alternative growth-area criteria assessed by day 28 after plating, including comparison with the day-14 measurement and a flask-area threshold of > 70%.

The registry also describes separate rapid-growth categories based on special staining: HCC cells alone, HCC cells with concomitant cancer-associated fibroblasts, or fibroblasts alone. The endpoint is used to examine whether a laboratory growth characteristic is associated with clinical stage. That makes it relevant to the study’s stated interest in primary culture as a potential research tool, but it does not directly measure treatment response or establish clinical utility.

The endpoint definition is reported, but the number of participants with rapid proliferation in each stage group, the effect estimate, and any measure of uncertainty are not reported in the registry record. Therefore, the direction and precision of the primary comparison cannot be determined from the listed endpoint information alone.

7. Why No Re-analysis Is Shown

8. Statistical Interpretation

9. Statistical Methods Explained

What does a binary endpoint represent?

General: A binary endpoint assigns each participant to one of two outcome states, such as meeting or not meeting a defined criterion. For this study, the primary outcome is whether a patient’s cultured cells satisfy the rapid-proliferation definition. The natural summary is a count and proportion in each stage group. These summaries preserve the participant as the unit of analysis and make clear how many observations contribute to each group. The registry identifies the endpoint unit as participants, but does not report the group counts or proportions.

How should a stage-group comparison be read?

General: A comparison of binary outcomes can be expressed as an absolute difference in proportions, a relative measure, or a model-based association. Each summary answers a different question. An absolute difference describes the gap between groups on the proportion scale, while a relative measure describes the ratio between them. A model may adjust for measured characteristics, but adjusted results depend on the variables and model specification. The registry record does not report the specific effect measure, statistical test, model, or adjustment variables used for the primary comparison. No additional effect measure can be inferred from the endpoint definition alone.

What does a confidence interval contribute?

General: A confidence interval presents a range of values compatible with the observed data under the interval’s statistical assumptions. It conveys uncertainty around an estimate; it is not a guarantee that the true value lies within the interval. For a comparison of proportions, the interval’s width is influenced by the amount of information in each group and by how common the outcome is. The registry record does not report a confidence interval for the primary endpoint, so the precision of the stage comparison cannot be assessed from the listed record details.

What assumptions matter for this endpoint?

General: Standard methods for comparing proportions rely on assumptions about how observations are sampled and how their outcomes relate to one another. In a participant-level comparison, the independence of observations is important. If observations are clustered, repeated, or otherwise correlated, ordinary methods can understate uncertainty unless the analysis accounts for that structure. The registry describes patients and cultured cells, but does not report whether the analysis accounted for clustering, repeated culture measurements, or other dependence. It also does not report the statistical method used to handle small or sparse group counts.

The rapid-growth rule combines two criteria based on growth area: comparison of measurements at day 14 and day 28, or a day-28 threshold of > 70% of flask area. General: When a composite classification can be met in more than one way, it is useful to understand the contribution of each component. Component-specific results can show whether the overall pattern is driven mainly by one criterion or is consistent across criteria. The registry describes the rule but does not report component-specific counts or results. It also does not report how ambiguous, unreadable, or unsuccessful cultures were classified for the primary comparison.

Why can over-dispersion matter?

General: Over-dispersion occurs when observed variability is greater than the statistical model expects. If a model assumes less variability than is present, its uncertainty estimates can be too narrow and significance tests can be overly optimistic. For binary outcomes, clustering or unmodelled differences among participants can contribute to over-dispersion in some settings. The registry does not report whether an over-dispersion assessment was relevant or performed, nor does it identify a model that would allow that question to be evaluated. This is a general consideration, not a claim that over-dispersion occurred in this study.

How should planning calculations be distinguished from study results?

General: A sample-size calculation is a planning exercise tied to assumptions about the effect to detect, allocation, significance level, and analysis method. It is not evidence that a study had a particular probability of detecting an effect after completion. The registry record’s planning illustration is described as based on the observed effect, with a two-sided significance level of 0.05, equal allocation between the two arms, and a normal approximation. For time-to-event outcomes, the stated planning convention uses total events; for other outcomes, it uses participants per arm. These are illustration assumptions, not reported design assumptions for the original prospective study. The original sample-size justification and its target effect are not reported in the registry record.

Because the primary comparison is between stage strata and not randomized treatment arms, equal-allocation planning assumptions should not be mistaken for the actual group sizes. The record does not report the number of participants in each stage category, and the planning convention does not establish that recruitment produced balanced groups. Any planning result is separate from the observed endpoint result, which is not reported in the registry record.

10. Multiplicity, Missing Data and Analysis Populations

The registry lists two outcome measures and nine analyses, but does not describe how those analyses relate to one another or which results were designated confirmatory. The primary outcome is identified, and a secondary outcome concerns worsening of AJCC TNM stage or HCC-related death 6 months after plating. The record does not report whether the analyses were prespecified in detail, whether adjustments for multiple comparisons were planned, or how any multiplicity was addressed. Without that information, the degree to which reported significance tests, if any, account for multiple testing cannot be assessed.

General: When many outcomes, subgroups, or analytic choices are examined, the chance of at least one apparently notable result can increase. Multiplicity procedures can change how evidence is interpreted, but their appropriateness depends on the planned questions and analysis strategy. The registry record does not provide enough detail to determine which procedures, if any, were used. Numerical findings and p-values for the listed outcomes are not reported in the registry record.

The number of participants with complete primary endpoint data, missing culture results, or exclusions from analysis is not reported in the registry record. Nor does it state how missing stage information, unsuccessful cultures, or incomplete follow-up were handled. The analysis population—such as whether all enrolled participants, only those with usable specimens, or another subset contributed to each analysis—is not reported in the registry record. These omissions limit assessment of possible selection effects and the amount of information available for each outcome.

11. What the Result Does, and Does Not, Mean

The registry describes a prospective investigation of culturing cells from residual fine-needle aspiration specimens and gives a rule for classifying rapid proliferation. It also specifies a comparison across AJCC TNM stage groups. These details explain what the investigators intended to evaluate, but they are not a report of the primary comparison’s observed result. The group counts, proportions, effect estimate, confidence interval, and p-value are not reported in the registry record.

General: An observed association between culture growth and stage would mean that the measured outcome differed across the groups in the study sample. It would not, by itself, show that stage caused the culture pattern, that the culture result predicts an individual patient’s future course, or that using the method improves treatment decisions. Those are separate questions that require appropriate evidence and analysis. The registry’s stated interest in potential personalized-treatment applications should therefore be distinguished from demonstrated clinical utility.

The secondary outcome refers to worsening stage or HCC-related death 6 months after plating, but its event counts and results are not reported in the registry record. The endpoint description alone cannot establish whether cultured-cell growth was associated with either later outcome. Similarly, the registry’s stated goal of evaluating culture success does not establish a success rate because the number of successful cultures is not reported.

The study enrolled 105 participants, but the number contributing usable data to the primary or secondary analyses is not reported in the registry record. Thus, the enrollment figure should not be treated as the analysis denominator. The reported completion status indicates the study is marked completed in the record; it does not fill the gap in outcome results. The appropriate interpretation is limited to the design and definitions recorded, with the study’s numerical findings and precision remaining unreported.

12. Why This Trial Matters Statistically

This study raises a statistically useful question about how a laboratory measurement might relate to a clinical classification. It defines a participant-level binary outcome from primary cell culture and compares that outcome across stage groups. Clearly defining the measurement rule matters because conclusions about an association depend on how growth is assessed, when it is measured, and what qualifies as rapid proliferation.

The distinction between an observational stage comparison and a randomized treatment comparison is also important. Stage was used to define the groups; it was not assigned as an intervention. Any difference between the groups would therefore need interpretation as an association, with attention to other participant, specimen, or laboratory factors that might influence culture growth. The registry record does not report whether those factors were measured or addressed analytically.

The study also connects a laboratory endpoint to a later clinical outcome, since its secondary outcome concerns worsening stage or HCC-related death 6 months after plating. That connection could motivate further research, but it requires reported event counts and estimates to evaluate. Those results are not reported in the registry record. The study’s stated interest in possible personalized-treatment applications is a rationale, not a finding that the method has clinical utility.

Finally, this record illustrates why endpoint definitions and numerical results need to be considered together. The growth criteria and stage categories are described, but the group counts, effect estimates, uncertainty, analysis population, and missing-data handling are not. Without those details, readers can understand the intended statistical question but cannot assess its answer or precision.

13. Limitations

• The posted summary statistics are the only data these results rest on; patient-level data, full protocols and statistical analysis plans are not available.
• Recalculated results are simple, unadjusted calculations from posted summaries. They do not replace the sponsor's prespecified analysis.

A central limitation for interpretation is that the registry record does not report the primary outcome values. It gives the rapid-proliferation definition and the stage comparison, but not the number of patients meeting the definition in either group, the resulting association estimate, or its uncertainty. The stated enrollment of 105 does not indicate how many participants had classifiable cultures or contributed to the primary analysis.

The primary outcome depends on culture growth after plating, yet the record does not report the number of attempted cultures, the culture success rate, or how unsuccessful and uninterpretable cultures were handled. It also does not give separate results for the day-14-to-day-28 growth criterion and the day-28 flask-area criterion. The distribution of rapid-growth categories based on special staining is not reported.

The stage groups are observational and may differ in characteristics beyond stage. The registry does not report adjustment methods, measured covariates, or whether any potential confounding was considered. It also does not report the analysis population, missing-data strategy, or whether the assumptions of the selected statistical method were assessed. The statistical method itself is not reported in the registry record.

For the secondary outcome, event counts, follow-up completeness, and analysis details are not reported in the registry record. The record also does not provide a sample-size justification for the original study or describe whether multiplicity was addressed across the listed analyses. These limitations prevent assessment of the strength, precision, and robustness of any reported association from the registry entries alone.

14. Related Tutorials

15. Related Statistical Calculators

16. Sources

• ClinicalTrials.gov: Primary Cell Culture of Hepatic Tumorous Cells From Routine Fine-needle Aspiration — NCT01549275.
• Registry record last updated 2015-10-08; status last verified 2013-07.
• Results first posted 2015-10-08.
• Data retrieved from ClinicalTrials.gov on 2026-11-09.
• Group sizes, means, event counts and sponsor-reported p-values are reproduced as posted. Effect estimates, confidence intervals and p-values not attributed to the sponsor are independently calculated from the posted summary statistics.

17. Record Summary

NCT01549275 is a completed prospective study of primary cell culture from residual specimens collected during routine fine-needle aspiration of hepatic tumors. The registry reports enrollment of 105 and lists hepatocellular carcinoma as the condition. The recorded study start is 2010-04 and primary completion is 2013-01.

The primary endpoint compares rapidly proliferative cultured cells between AJCC TNM stage IIIA or lower and stage IIIB or higher, using growth-area criteria assessed at 28 days after plating. A secondary outcome concerns worsening stage or HCC-related death 6 months after plating. The record lists two outcome measures and nine analyses, but does not report the numerical endpoint results, analysis methods, or analysis populations.

The registry identifies Kaohsiung Medical University Chung-Ho Memorial Hospital as sponsor and names Kaohsiung Medical University Hospital in the official title field. The study’s stated purpose included evaluating culture success and considering possible personalized-treatment applications; the culture success rate and clinical application results are not reported in the registry record.