Culturing and robotic harvesting of bone marrow, lymph nodes, peripheral blood, fibroblasts, and solid tumors with in situ techniques

Spurbeck, Jack L.; Carlson, Richard O.; Allen, Judith E.; Dewald, Gordon W.

doi:10.1016/0165-4608(88)90312-3

Cited by 71 publications

(22 citation statements)

References 3 publications

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“…The cultures were harvested using a Tecan robotic harvest system MiniPrep75™ using the standard method for in situ cultures. 27 The coverslips were dried in a Thermotron Slide Drying Chamber CDS-5™. 28 After slide drying, the slides were submerged in a trypsin working solution followed by a water rinse.…”

Section: Specimen Preparationmentioning

confidence: 99%

Reflex fluorescent in situ hybridization testing for unsuccessful product of conception cultures: A retrospective analysis of 5555 samples attempted by conventional cytogenetics and fluorescent in situ hybridization

Shearer

Thorland

Carlson

et al. 2011

Genetics in Medicine

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Introduction:The use of chromosome analysis on products of conception from spontaneous abortions is recommended to identify a genetic etiology. However, 20% of products of conception cultures are unsuccessful due to microbial contamination or lack of viable dividing cells. Our laboratory implemented a reflex fluorescent in situ hybridization (FISH) assay to detect numeric chromosome abnormalities for unsuccessful cultures. Materials and Methods: All products of conception samples were simultaneously processed for both chromosome analysis and FISH analysis. If the chromosome analysis was unsuccessful, interphase FISH was performed for chromosomes 13,16,18,21,22, X, and Y. To assess the performance of the FISH assay, a 3-year retrospective comparative analysis of the FISH results versus chromosome results was performed. Results: Of 5555 total specimens, 4189 (75%) represented chorionic villi/fetal tissue and 1366 (25%) represented tissue of unidentified origin. Of the 1189 tissues of unidentified origin with chromosome or FISH results, 1096 (92%) were XX, indicating that the majority of these tissues are likely maternal in origin. Of the 3361 successful chromosome studies on the chorionic villi/fetal tissue specimens, 1734 (52%) samples had a chromosome abnormality. Of the 762 successful FISH studies on chorionic villi/fetal tissue specimens that were unsuccessful by chromosome studies, 181 (25%) had an abnormal result with the targeted FISH panel. Overall, the FISH panel detected approximately 70% of the chromosome abnormalities in products of conception detectable by karyotype. When the FISH panel results were combined with chromosome analysis for the 4189 chorionic villi/fetal tissue specimens, the overall abnormality rate is 47%. Conclusions: Our reflex FISH assay proved useful for the detection of common chromosome aneuploidies in products of conception samples that failed conventional chromosome analysis. Because of its limited view of the genome, cautious interpretation of FISH results is required for all samples, in particular, trisomy of an acrocentric chromosome, which may represent a Robertsonian translocation. An algorithmic approach to the genetic evaluation of products of conception specimens, with the potential for initial evaluation by a FISH panel, may be warranted. Genet Med 2011:13(6):545-552.

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Section: Specimen Preparationmentioning

confidence: 99%

Reflex fluorescent in situ hybridization testing for unsuccessful product of conception cultures: A retrospective analysis of 5555 samples attempted by conventional cytogenetics and fluorescent in situ hybridization

Shearer

Thorland

Carlson

et al. 2011

Genetics in Medicine

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“…For cytogenetic analysis, bone marrow specimens were processed by both direct and short-term culture techniques as described previously. 14,15 Circulating plasma cells were estimated based on a similar method as described elsewhere. 16 Circulating plasma cells were defined as a poor prognostic factor when у3% of cytoplasmic immunoglobulinpositive cells were considered plasma cells by morphologic examination.…”

Section: Prognostic Factors and Risk Groupsmentioning

confidence: 99%

Effect of complete response on outcome following autologous stem cell transplantation for myeloma

Rajkumar

Fonseca

Dispenzieri

et al. 2000

Bone Marrow Transplant

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Summary:We studied the effect of complete response (CR) among 126 consecutive patients who underwent stem cell transplantation (SCT) for myeloma. The CR rate with SCT was 33%. Median overall survival (OS) from diagnosis of myeloma was 56 months. OS following SCT was 22 months. Progression-free survival (PFS) was 12 months. OS was not different between patients who achieved CR and those who did not, median survival 25 vs 24 months, P = 0.5. Corresponding median times for PFS were 15 and 11 months, P = 0.2. The plasma cell labeling index (PCLI) was high (у1%) in 36% (high risk group) and was associated with poor OS and PFS (P Ͻ 0.001). Achieving CR did not influence OS or PFS in either the high or the low risk group. In contrast, OS and PFS were significantly influenced by high PCLI both in patients who achieved CR and those who did not. OS was poor (Ͻ30 months) in high risk patients regardless of CR status and in low risk patients who did not achieve CR, compared to low risk patients achieving CR (57 months), making them candidates for novel posttransplant treatment options. Outcome following SCT is dependent more on biological variables such as the PCLI than on CR status. Bone Marrow Transplantation (2000) 26, 979-983. Keywords: multiple myeloma; complete response; prognosis; transplantation Multiple myeloma is a clonal plasma cell proliferative disorder that is characterized by the presence of a monoclonal (M) protein in the serum or urine, lytic bony lesions, renal failure, anemia and hypercalcemia. It accounts for 1% of all cancers, and 10% of all hematologic malignancies.1,2 In 2000, approximately 13 700 new cases of myeloma will be diagnosed and over 11 000 will die of the disease in the United States.2 Therapy for myeloma is not curative. The overall response rate with standard melphalan and prednisone therapy is approximately 50% and the complete response (CR) rate is less than 10%.3,4 Combination chemotherapy leads to superior response rates (60-70%), but is without survival benefit.3,5,6 High-dose therapy followed by autologous blood cell transplantation results in response rates exceeding 75-90%, 4,5 and CR rates in the range of 20-40%. 7,8 A French randomized trial has demonstrated improved survival with autologous marrow transplantation compared to conventional chemotherapy, with 5 year survival rates of 52% and 12%, respectively.

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“…For cytogenetic analysis, bone marrow specimens were processed by both direct and short-term culture techniques, as described previously. 13,14 The 2 and Fisher exact tests were used to compare differences among the three subgroups for nominal variables; the rank-sum test was used for continuous variables. Survival analysis was done using the method described by Kaplan and Meier.…”

Section: Approval Of the Mayo Foundation Institutional Reviewmentioning

confidence: 99%

Autologous stem cell transplantation for relapsed and primary refractory myeloma

Rajkumar

Fonseca

Lacy

et al. 1999

Bone Marrow Transplant

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Summary:The aim of this study was to evaluate the effectiveness of autologous stem cell transplantation for patients with relapsed or primary chemotherapy-refractory myeloma. Seventy-five patients, 50 men and 25 women, ages 33-68 years (median, 53 years), who underwent transplantation for relapsed or primary refractory myeloma were studied. Patients underwent transplantation 5-88 months (median, 23 months) after diagnosis of myeloma. The time to transplantation was significantly shorter in patients with refractory disease than for those with relapsed myeloma (median, 8 and 32 months, respectively; P Ͻ 0.001). Patients with primary refractory myeloma had a significantly lower plasma cell labeling index than those with relapsed disease (P ‫؍‬ 0.008). There were no differences in overall and complete response rates between patients with primary refractory and relapsed disease. The median survival of the entire cohort from diagnosis was 53 months. Overall survival from transplantation among patients with relapsed myeloma receiving therapy, relapsed myeloma off therapy, and primary refractory myeloma was significantly different (P ‫؍‬ 0.04), with median times of 12, 21 and 30 months, respectively. Progression-free survival also was different (P Ͻ 0.001), with median times of 7, 13, and 26 months, respectively. We conclude that overall and progression-free survival in patients with primary refractory myeloma appear better than in patients with relapsed disease and need further study.

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Culturing and robotic harvesting of bone marrow, lymph nodes, peripheral blood, fibroblasts, and solid tumors with in situ techniques

Cited by 71 publications

References 3 publications

Reflex fluorescent in situ hybridization testing for unsuccessful product of conception cultures: A retrospective analysis of 5555 samples attempted by conventional cytogenetics and fluorescent in situ hybridization

Reflex fluorescent in situ hybridization testing for unsuccessful product of conception cultures: A retrospective analysis of 5555 samples attempted by conventional cytogenetics and fluorescent in situ hybridization

Effect of complete response on outcome following autologous stem cell transplantation for myeloma

Autologous stem cell transplantation for relapsed and primary refractory myeloma

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