Organotial Human hepatobiliary carcinoma organoid Culture Kit
SKU: 42614884356

Organotial Human hepatobiliary carcinoma organoid Culture Kit

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Description

Organotial Human hepatobiliary carcinoma organoid Culture KitProduct Specification Description Component name Specification Human hepatic bile duct carcinoma organoid medium A 100mL Primary culture buffer B 250mL Human hepatic bile duct carcinoma primary tissue digestive juice C 30mL Organoid passage digestive juice D 30mL Tissue preservation solution E 100mL Organoid cryopreservation fluid F 20mL Organoid subculture buffer G 250mL Instructions 1, primary (1) The harvested tissue should be placed in precooled

Product Specification

Description
Component name Specification
Human hepatic bile duct carcinoma organoid medium A 100mL
Primary culture buffer B 250mL
Human hepatic bile duct carcinoma  primary tissue digestive juice C 30mL
Organoid passage digestive juice D 30mL
Tissue preservation solution E 100mL
Organoid cryopreservation fluid F 20mL
Organoid subculture buffer G 250mL
Instructions 1, primary
(1) The harvested tissue should be placed in precooled (2-8° C) In the sampling bottle of tissue preservation solution E, the tissue was quickly transferred to a clean laboratory for tissue processing and cell separation, photographed and recorded.
(2) Prepare several culture dishes and add primary culture buffer B precooled at 4 ° C for later use.
(3) The sampling bottle was disinfected, and the tissue was placed in the Petri dish. After washing three times with primary culture buffer B, the tissue was cut into tissue blocks of about 1-3mm3in volume using ophthalmic scissors or scalpel.
(4) The tissues were digested with human hepatobiliary carcinoma primary tissue digestion solution C and shaken at 37℃ for 15-25min (the digestion was observed at any time during the digestion).
(5) A small amount of liquid was observed under the microscope. When more single cells or cell clusters below 70um were observed under the microscope, triploid volume primary culture buffer B was added to terminate the digestion.
(6) The filtrate was filtered by a sieve with a pore size of 100um, and the filtrate was collected. After enrichment and centrifugation at 300g for 5min, the supernatant was removed, and the primary culture buffer B was added and resuspended for centrifugation.
(7) Matrix glue calculation: The volume of collected tissue was observed after step 6, and matrix glue (abs9495) was added 25 times the tissue volume to resuspend the paving plate.
(8) For the example of 24-well cell culture plates, 25uL of tissue matrix glue mixture was dispensed to each well for paving (operation at 4 ° C).
(9) The paved culture plate was placed in an incubator at 37 ° C for 10-15min to form gelatin, and then cultured with human hepatobiliary carcinoma organoid medium A (restored to room temperature).
2, Organoid subculture
(1) The medium was sucked out by pipetted gun, and 1-2 ml of 4℃ organoid subculture buffer G was added to each well for 2min.
(2) The matrix glue was gently blown by a pipetting gun, collected in a 15mL centrifuge tube and allowed to rest for 10min at 4 ° C. (every 6-8 Wells were a group)
(3) a: When the number of organoids was insufficient or the volume was small: the supernatant was discarded by centrifugation for 5min, then an appropriate amount of organoid passage culture buffer G was added and resuspended into a 1.5mL centrifuge tube, and the liquid was discarded by centrifugation at 300g for 5min for the fourth step.
          b: When organoids are large in number or size: The supernatant was discarded by centrifugation for 5min, then the organoid passage digestion solution D was added for 2-3min, then the organoid passage culture buffer G was added to terminate the digestion, the mixture was discarded by centrifugation for 5min, then the organoid passage culture buffer G was added and the mixture was resuspended and transferred to a 1.5mL centrifuge tube, and the liquid was discarded by centrifugation at 300g for 5min for step 4.
(4) After the organoids were collected, 25uL of matrigel was added to the 24-well cell culture plate, and 500uL of human hepatobiliar carcinoma organoid medium A was added to the incubator for 10-15min.
3. Organoids were frozen
(1) The medium was removed by suction with a pipettes gun, and 1-2 ml of 4 ° C organoid passage culture buffer G was added to each well and placed for 2min.
(2) The matrix glue was gently blown by a pipetting gun, collected in a 15mL centrifuge tube and allowed to rest for 10min at 4 ° C. (every 6-8 Wells were a group)
(3) The supernatant was discarded by centrifugation for 5min, then an appropriate amount of organoid passage culture buffer G was added and resuspended again, and the liquid was discarded by centrifugation at 300g for 5min.
(4) Add appropriate amount of organoid cryopreservation solution F, gently blow and resuspend, take 24-well cell culture plate as an example: the density is 2 Wells and freeze 1 tube, each tube volume is 1.4mL.
(5) After making the labeling information and cooling, the samples were transferred to liquid nitrogen for long-term storage.
4, organoid resuscitation
(1) 10mL organoid subculture buffer G was placed in a 15mL centrifuge tube.
(2) The frozen organoid cells were removed from the liquid nitrogen tank and quickly thawed in a 37℃ water bath.
(3) During the water bath melting process, it is necessary to gently shake the freezing tube to ensure that the frozen solution is completely melted within 1-2min.
(4) The dissolved organoid cells were quickly transferred to a 15mL centrifuge tube, gently blown with a pipette for 6-8 times, centrifuged at 300g for 5min, and then the supernatant was removed and the organoid cell precipitate was collected. An appropriate amount of organoid passage culture buffer G was added and resuspended into a 1.5mL centrifuge tube and centrifuged at 300g for 5min.
(5) Matrigel was resuspended, 25uL of Matrigel per well was spread in A 24-well cell culture plate, placed in an incubator for 10-15min to gel, and 500uL of human hepatobiotic carcinoma organoid medium A was added.
Storage Temp. Stored at 4 ° C, valid for 3 months; Stored at -20 ° C, valid for 1 year, see label for details.
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SKU: 42614884356

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4.6 ★★★★★
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Amanda Greathouse
West Palm Beach, US
★★★★★ 3
3.5 stars, A little boring to say the least.
Format: Kindle
Wow so I'm not sure where to begin on this one. This was a very different take on the legend of Arthur and Excalibur. This is told from the point of view of Morgan the sister of Arthur. Honestly the first 50% of this book is world building and character building which unfortunately was super boring for me. Morgan to me was a female MC that had a hard time in believing in herself. Sometimes taking too long to understand exactly what was going on around her. Draven was also a different male MC, like I couldn't put my finger on him and what he was all about. It was not until the last 10% of the book did we get some answers on the mystery that is Draven. The other 50% of the book centered around this big journey with everyone having a different motive. We see a spark of magic around this time that had me excited but then we never expanded upon that and what it could mean for the female MC. I feel like I want to read the second book just to see where this goes, but the spice was probably a 2 out of 5. Side characters are ok, Lancelet was fun but I almost felt like I wanted more.
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Reviewed in the United States on September 13, 2023
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Krystina
Belleville, US
★★★★★ 5
A bewitching retelling of Arthurian legend!
Format: Kindle
In a land where the Fae have nearly become only a legend and those who still posses even a morsel of the blood are few and far between, Morgan finds herself cast aside by most of society due to her rumored half-Fae lineage, including her brother, King Arthur. With the kingdom at the brink of war, Arthur entrusts her with a quest to retrieve a Fae weapon of legendary power: the sword of Perun, Excalibur. Accompanied by men she loathes, Captain Kairos Draven and Ragnar Whitehorn, she embarks on her long and unbeknownst perilous journey, only to find that things she once believed to be myth are in fact very real. With devastating twists, omitted truths, witty banter and fierce action, Queen of Roses leaves you begging to know more about the secrets of Aercanum! Wow, wow, wow! Going into this story, I did not realize that it was going to be a retelling of Arthurian legend, especially not one with a fantastical twist! The unique spin almost gave me The Witcher vibes and I think adding Fae into the mix was quite interesting. I knew the basics of the legend but after reading this book, it has piqued my interest and makes me want to learn more about it. My attention was snatched as soon as I finished the prologue and I knew that I was going to devour this story. I truly enjoyed the gender swaps and even how Arthur was portrayed as villainous. Morgan’s past and even her parts of her present is absolutely heartbreaking, and I felt for her at times. I can only recall one other book that made me hate characters the way I despised Florian and Arthur, leaving me with my blood boiling and feeling disgusted. Even after finishing the book, Draven is still a mystery to me and I cannot figure out how to feel about him. I guess they just means that the author did an excellent job at conveying each character’s persona! The rich world building and imagery made it easy for me to visualize the places that the group visited along their journey. I am truly engulfed in this story and I cannot wait to see wait fate awaits Morgan and how the Fae will be even more incorporated in the next book!. I received a free copy of this book and am voluntarily leaving a review.
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Reviewed in the United States on August 11, 2023
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Tiana
Massapequa, US
★★★★★ 4
Enchanting
Format: Kindle
"Queen of Roses" by Briar Boleyn is a delightful and refreshing reimagining of the classic tale of King Arthur, with a captivating twist that places the spotlight on Morgan, a character who has often been overshadowed in traditional retellings. Boleyn's creative decision to shift the narrative perspective to Morgan breathes new life into the story, offering readers an intriguing and compelling look at the Arthurian world from an entirely different angle. One of the most commendable aspects of this book is its incorporation of Fae elements, which adds an enchanting layer of magic and mystery to the already familiar Arthurian setting. Boleyn skillfully weaves the world of the Fae into the narrative, creating a captivating backdrop against which the events of the story unfold. This addition not only adds depth to the world-building but also provides ample opportunities for twists and turns that keep readers thoroughly engrossed. However, while the book boasts numerous strengths, it does have one noticeable flaw: the characterization of Morgan. While it is reasonable to create a flawed and complex protagonist, it appears that at times, Morgan's character becomes overly difficult and hard to relate to. Her persistently negative perception of one of the main male characters, who is a potential love interest, despite his efforts to support and assist her, may come across as somewhat irrational and could test the patience of some readers. Striking a balance between a strong, independent character and one who can recognize genuine support and affection could have enhanced the overall reader experience. Nonetheless, the allure of "Queen of Roses" lies in its innovative approach to the Arthurian legend and its skillful blending of fantasy elements into a familiar narrative. Boleyn's evocative prose draws readers into a world where magic, destiny, and fate entwine, leaving us eager to uncover the mysteries that unfold within the pages. I received a free copy of this book via Booksprout and am voluntarily leaving a review.
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Reviewed in the United States on July 28, 2023
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Stephanie
Fort Morgan, US
★★★★★ 5
An action-packed dark romantasy
Format: Kindle
I loved this book! Queen of Roses is an Arthurian-inspired dark romantasy that is the first book in the Blood of Fae series. The story follows Morgan, the princess of Camelot who is rumored to be part fae. Fueled by prejudiced hatred and a mistrust of fae blood, Morgan’s abusive father strips her of her birthright and hands it to her half-brother, Arthur. Instead of becoming queen, Morgan is commanded to join the temple of the goddesses when she comes of age. However, Arthur turns into a psychopathic, power-hungry, fae-hating king as he ages. He develops malevolent plans and commands Morgan to find an ancient weapon with legendary power. Although Morgan is wary of Arthur’s intentions, she embraces the opportunity to go on a journey and potentially change her fate. The story picks up from there and we follow Morgan on her quest to find the ancient relic. It’s full of high stakes adventure, mystery, tension, banter, forced proximity, hidden magic, self discovery, and betrayal. This first installment of the series intricately develops the world building and character development. There’s little romance in this book, but it is evident that it is a slow burn that will continue to develop throughout the remainder of the series. Overall, I loved the world building, the epic fantasy, Morgan’s journey of self discovery, and all of the twists and turns that set the stage for the future installments. I can’t wait to see what happens next!
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Reviewed in the United States on April 7, 2024
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AlynReads
Omaha, US
★★★★★ 4
Arthurian Fae Quest…say less.
Format: Kindle
A fae centered Arthurian tale unlike any I’ve read so far. The author did a great job at descriptive world building, with scenes easily playing out in my minds eye. There was plenty of action, suspense, and even a touch of horror. An enemies to lovers, slow burn romance, a quest, with plot twist and turns aplenty. There was a love triangle, which I’m not usually a fan of but, it played out well in this story line. The FMC, Morgan Pendragon, was so blatantly naïve, yet I typically expect as much in a ‘book one’ of a series, especially one that features a fairly sheltered princess. I was happy to read that in spite of this, she still showed a strong sense of morals, fire, and spine. Now our MMC? Kairos Draven, aka Void’s Edge. Oh, how I’m a sucker for a smoking’ hot grumpy warrior alpha with a witty mouth, and a strong sense of “touch her and die” attitude, so you know who held all my cards. That ending? Just made me swoon all the harder. Now add a battlecat that rivals the size of a horse…and well Ms. Briar Boleyn you have well and truly stolen my heart. I’m excited to see where the story goes from here, and follow along to see more of the characters growth. I went into this story fairly blind, and I think I enjoyed it all the more because of it. Once the story got going, it had me in an absolute chokehold and it was difficult to put down.
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Reviewed in the United States on May 12, 2024

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